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# Helix Formation and Folding in an Artificial Peptide

## I Introduction

The mechanism by which a large class of proteins folds spontaneously into a unique globular shape  has remained elusive. Significant new insight was gained over the last few years from the studies of minimal protein models. For instance, energy landscape theory and funnel concept  proved to be powerful tools for description of the general characteristics of folding not only in minimalistic protein models but also for real proteins . However, many questions on the details of the folding process remain to be solved. For instance, folding of proteins involves one or more transitions between different thermodynamic states. The role of these transitions in the folding process is an active area of research. An important example for these transitions is the formation of secondary structure elements. For the case of $`\alpha `$-helices it is long known that there is a sharp transition towards a random coil state when the temperature is increased. The characteristics of this so-called helix-coil transition have been studied extensively , most recently in Refs. . In this paper, we research the relation between helix-coil transition and folding.

For this purpose, we have studied an artificial peptide, Ala10-Gly5-Ala10, in a detailed representation where the interactions between all atoms are taken into account. Multicanonical simulations  with large statistics are used to evaluate the free energy landscape of our peptide at different temperatures. The encountered transitions are further investigated by partition zeros analysis which allows to characterize “phase transitions” in small systems . Quantities such as the energy, specific heat, helicity and susceptibility were calculated as function of temperature. We have neglected in the simulations the interaction of our artifical peptide with the surrounding solvent. While this is certainly a crude approximation, it allows us not only to relate our results to our previous studies on helix-coil transition in poly-alanine that also relied on gas-phase simulations , but also to study the extend to that secondary structure formation and folding are determined by intrinsic properties of the peptide. Our data suggest that the peptide in gas-phase folds in a two-step process: in a first step two $`\alpha `$-helices are formed in what amounts to a first order transition. Afterwards these helices re-arrange themselves into a U-like structure. The second step has the characteristics of a second order transition.

## II Methods

Our investigation of Ala10-Gly5-Ala10 is based on a detailed, all-atom representation of that peptide. The interaction between the atoms is described by a standard force field, ECEPP/2, (as implemented in the program package SMMP ) and is given by:

$`E_{tot}`$ $`=`$ $`E_C+E_{LJ}+E_{HB}+E_{tor},`$ (1)
$`E_C`$ $`=`$ $`{\displaystyle \underset{(i,j)}{}}{\displaystyle \frac{332q_iq_j}{ϵr_{ij}}},`$ (2)
$`E_{LJ}`$ $`=`$ $`{\displaystyle \underset{(i,j)}{}}\left({\displaystyle \frac{A_{ij}}{r_{ij}^{12}}}{\displaystyle \frac{B_{ij}}{r_{ij}^6}}\right),`$ (3)
$`E_{HB}`$ $`=`$ $`{\displaystyle \underset{(i,j)}{}}\left({\displaystyle \frac{C_{ij}}{r_{ij}^{12}}}{\displaystyle \frac{D_{ij}}{r_{ij}^{10}}}\right),`$ (4)
$`E_{tor}`$ $`=`$ $`{\displaystyle \underset{l}{}}U_l\left(1\pm \mathrm{cos}(n_l\chi _l)\right).`$ (5)
Here, $`r_{ij}`$ (in Å) is the distance between the atoms $`i`$ and $`j`$, and $`\chi _l`$ is the $`l`$-th torsion angle. The peptide bond angles were set to their common value $`\omega =180^{}`$. We do not include explicitly the interaction of the peptide with the solvent into our simulations and set the dielectric constant $`ϵ`$ equal to 2. Since the charges at peptide termini are known to reduce helix content , we removed them by taking a neutral NH2– group at the N-terminus and a neutral –COOH group at the C-terminus.

Simulation of detailed protein models where the interaction between all atoms are taken into account are extremely difficult. This is because the various competing interactions within the molecule lead to an energy landscape characterized by a multitude of local minima separated by high energy barriers. Hence, in the low-temperature region, canonical Monte Carlo or molecular dynamics simulations will tend to get trapped in one of these minima and the simulation will not thermalize within the available CPU time. One example of the new and sophisticated algorithms  that allow to overcome this difficulty are generalized-ensemble techniques , and it is one of these techniques, multicanonical sampling , that we used for our investigations.

In the multicanonical algorithm  conformations with energy $`E`$ are assigned a weight $`w_{mu}(E)1/n(E)`$. Here, $`n(E)`$ is the density of states. A simulation with this weight will lead to a uniform distribution of energy:

$$P_{mu}(E)n(E)w_{mu}(E)=\mathrm{const}.$$
(6)
This is because the simulation generates a 1D random walk in the energy space, allowing itself to escape from any local minimum. Since a large range of energies are sampled, one can use the reweighting techniques  to calculate thermodynamic quantities over a wide range of temperatures $`T`$ by

$$<𝒜>_T=\frac{𝑑x𝒜(x)w^1(E(x))e^{\beta E(x)}}{𝑑xw^1(E(x))e^{\beta E(x)}},$$
(7)
where $`x`$ stands for configurations.

Note that unlike in the case of canonical simulations the weights

$$w(E)=n^1(E)=e^{S(E)}$$
(8)
are not a priori known. Instead estimators for these weights have to be determined by an iterative procedure . In our case we needed 500,000 sweeps for the weight factor calculations. All thermodynamic quantities were then estimated from one production run of $`8,000,000`$ Monte Carlo sweeps which followed 10,000 sweeps for thermalization. Our simulations were started from completely random initial conformations (Hot Start) and one Monte Carlo sweep updates every torsion angle of the peptide once. At the end of every 10th sweep we stored the ECEPP/2 energies $`E_{tot},E_C,E_{LJ},E_{hb}`$ and $`E_{tor}`$ of the conformation, the corresponding number $`n_H`$ of helical residues and end-to-end distance $`d_{ee}`$. Here, we follow previous work  and consider a residue as helical if its backbone angle $`(\varphi ,\psi )`$ are within the range $`(70^{}\pm 20^{},37^{}\pm 20^{})`$.

Using the results of our generalized-ensemble simulation, we explored for various temperatures the free energies

$$G(n_H,d_{ee})=k_BT\mathrm{log}P(n_H,d_{ee}).$$
(9)
Here, $`P(n_H,d_{ee})`$ is the probability to find a peptide conformation with values $`\mathrm{}`$, $`d_{ee}`$ (at temperature $`T`$). We chose the normalization so that the lowest value of $`G(n_H,d_{ee})`$ is set to zero for each temperature.

We finally used that the multicanonical algorithm allows us to calculate estimates for the spectral density:

$$n(E)=P_{mu}(E)w_{mu}^1(E).$$
(10)
We can therefore construct the corresponding partition function for our all-atom model of Ala10-Gly5-Ala10 from these estimates by

$$Z(\beta )=\underset{E}{}n(E)e^{\beta E},$$
(11)
with $`\beta `$ the inverse temperature, $`\beta =1/k_BT`$. The complex solutions of the partition function determine the critical behavior of the model and were also studied by us.

## III Results and Discussion

Our peptide, Ala10-Gly5-Ala10, is build up out of two chains of each 10 alanine residues connected by 5 glycine residues. In previous work  we could show that polyalanine has a pronounced transition between a disordered coil phase and an ordered state in which the polymer forms an $`\alpha `$-helix. For this reason, we expect formation of $`\alpha `$-helices in our peptide, and the average number of helical residues $`<n_H>`$ is therefore one of the quantities that we have measured. $`<n_H>`$ is displayed in Fig. 1 as a function of temperature, and we observe in this plot two temperature regions. At high temperature, few residues are found with backbone dihedral angles $`(\varphi ,\psi )`$ typical for an $`\alpha `$-helix. On the other hand, at low temperatures we observe helix-formation, and almost all of the alanine residues are part of an $`\alpha `$-helix, i.e. have backbone dihedral angles $`(\varphi ,\psi )`$ in the range ($`70^{}\pm 20^{},37^{}\pm 20^{}`$). The transition between the two temperature regions is sharp indicating the existence of a helix-coil transition. The transition temperature $`T_{hc}`$ can be determined from the corresponding peak in the susceptibility

$$\chi (T)=<n_H^2(T)><n_H(T)>^2,$$
(12)
which is ploted in the inset of Fig. 1, and we find the transition temperature $`T_{hc}=485\pm 5`$ K.

In previous work  we could show that in polyalanine the formation of $`\alpha `$-helices is related to a gain in potential energy. For this reason, we display in Fig. 2 the average total ECEPP/2 energy $`<E_{tot}>`$ and the thermodynamic averages of partial energies $`<E_C>`$, $`<E_{LJ}>`$, $`<E_{hb}>`$ and $`<E_{tor}>`$ of Ala10-Gly5-Ala10 as a function of temperature. As expected, we observe around our transition temperature $`T_{hc}`$ a sharp decrease in the potential energy $`<E_{tot}>`$ that is due to a corresponding decrease in the Coulomb energy $`<E_C>`$, Lennard-Jones energy $`<E_{LJ}>`$ and hydrogen-bond energy $`<E_{hb}>`$. The change in $`<E_{tot}>`$ with temperature can be described by the specific heat

$$C(T)=\beta ^2\frac{<E_{tot}^2><E_{tot}>^2}{25},$$
(13)
which we display in Fig. 3. A pronounced change in energy with temperature corresponds to a peak in the specific heat. As one can see from Fig. 3, we observe indeed a pronounced peak in the specific heat at a temperature $`T=480\pm 10`$ K that is consistent with $`T=485\pm 5`$ K, the temperature where the peak in the susceptibility is located. Combining both values we obtain as our final estimate for the helix-coil transition temperature $`T_{hc}=483\pm 8`$. However, we find in Fig. 3 also a second, smaller peak in the specific heat at the lower temperature $`T_f=265\pm 7`$ K indicating yet another transition.

The sharp decrease in potential energy corresponding to this second peak is clearly visible in Fig. 2 for the total energy $`<E_{tot}>`$, however, not for all of the partial energies. Only the Lennard-Jones term $`<E_{LJ}>`$ exhibits also a signal for the second transition at $`T_f`$. This energy term depends strongly on the overall size of the molecule and the change in this quantity indicates a transition between extended and compact structures. Hence, we conjecture that the second peak in specific heat at the lower temperature $`T_f`$ is related to a transition between extended and compact structures. A possible measure for such a change is the average end-to-end distance $`<d_{ee}>_T`$. We define here $`<d_{ee}>`$ as the distance between N of Ala1 and O of Ala25, and plot this quantity in Fig. 4. We observe that this quantity decreases with decreasing temperature. Below the helix-coil transition $`T_{hc}`$ the decrease slows down and the curve becomes almost flat at a value of $`<d_{ee}>10`$ Å indicating that there is little further change in the compactness of the molecule. However, at temperature $`T_f`$ the end-to-end distance decreases again sharply towards a new value $`<d_{ee}>=6.1`$ Å. Hence, $`T_f`$ marks the folding of the molecule into a defined compact structure with the two terminal ends of the peptide close together.

The transition between extended and a more compact structure can also be seen when we display the free energy landscape of our peptide as a function of helicity $`n_H`$ and end-to-end distance $`d_{ee}`$. At the temperature $`T=480`$ K (which is essentially the helix-coil transition temperature $`T_{hc}=483\pm 8`$ K) the free energy landscape (displayed in Fig. 5a) is flat over a large range of values of $`n_H`$ and $`d_{ee}`$. The $`3k_BT`$ contour line surrounds a region where the helicity can take values between $`0n_H20`$ and the end-to-end distance values between $`3d_{ee}40`$, allowing the system to move freely between extended and compact configurations, and between helical and coil configurations. On the other hand, at the second and lower temperature $`T=270`$ K (which is essentially the folding transition temperature $`T_f=265\pm 7`$ K) the free energy grows rapidly with decreasing helicity $`n_H`$ favoring configuration in a small strip with $`15n_H25`$. Hence, the plot of the free energy landscape in Fig. 5b is limited to values $`12n_H25`$ of the helicity. Here, two regions of minimal free energy can be seen (marked by the 3 $`k_BT`$ contour lines). The first minima is found at values of $`d_{ee}`$ between 5 and 10 Å and $`15n_H20`$ characterising compact structures. A second region with slightly lower free energy (see the 1 $`k_BT`$ contour line) is found at much larger values of $`d_{ee}`$ between 35 and 40 Å and $`20<n_H25`$ indicating a long stretched $`\alpha `$-helix. Both local free energy minima are separated by free energy barriers of $`8`$ $`k_BT`$ that can be overcome by thermal fluctuations. On the other hand, configurations with helicity $`n_H<10`$ are suppressed by free energy differences of more than 30 $`k_BT`$.

Examples for the structures corresponding to the two free energy minima are plotted in Fig. 6. The first one, displayed in Fig. 6a is the configuration with lowest energy ever found in our multicanonical simulation of 8,000,000 sweeps and corresponds to the region in the free-energy landscape at values of $`d_{ee}`$ between 5 and 10 Å and $`15n_H20`$. This conformation (‘A’) consists out of two helixes (made up out of the alanine residues) connected by a turn (build out of the flexible glycine residues) towards a U-turn-like structure that is consistent with the small value of the end-to-end distance $`d_{ee}`$ observed in Fig. 4 for temperatures below $`T_f`$. For reference we show in Fig. 6b also the configuration (‘B’) where all 25 residues are part of an $`\alpha `$-helix and which corresponds to the second local free-energy minimum in Fig. 5b at values of $`d_{ee}`$ between 35 and 40 Å and $`20<n_H25`$ . The dihedral angles of both configurations are listed in Table 1. Fig. 7 displays the frequency of both configurations as a function of temperature. For $`T>T_{hc}`$ neither configuration ‘A’ nor ‘B’ are observed. Below that temperature both structures appear with similar probability as long as the temperature is higher than $`T_f`$. At $`T=T_f`$ the probability to find the maximal helical structure ‘B’ is with $`30`$ % highest and decreases after that with decreasing temperature. On the other hand, the frequency for the U-turn structure ‘A’ continues to grow with decreasing temperature. This different behavior is due to the energy differences between both structures. The minimal energy conformation ‘A’ has with $`E_{tot}=34.7`$ Kcal/mol a $`10.8`$ Kcal/mol lower potential energy than the extended helix conformation ‘B’ ($`E_{tot}=23.9`$ Kcal/mol). This difference is mainly due to the Lennard-Jones terms: $`E_{LJ}=132.5`$ Kcal/mol for ‘A’ vs. $`E_{LJ}=118.9`$ Kcal/mol for ‘B’. The gain in $`E_{LJ}`$ is in part compensated by the hydrogen-bonding terms: $`E_{hb}=30.2`$ Kcal/mol for ‘A’ vs. $`E_{hb}=34.7`$ Kcal/mol for ‘B’. Coulomb and torsion energies differed little between the two configurations: $`E_C=126.7`$ Kcal/mol and $`E_{tor}=1.3`$ Kcal/mol for conformation ‘A’ vs. $`E_C=126.4`$ Kcal/mol and $`E_{tor}=3.3`$ Kcal/mol for conformation ‘B’.

It is an interesting question whether our two observed transitions (occuring in a finite and small system) can be related to phase transitions which in a strict sense are defined only for macroscopic (that is very large) systems. In order to study this question we have calculated the complex zeros $`\beta \mathrm{Re}(\beta )+i\tau `$ of the partition function $`Z(\beta )`$ of our molecule. In the case of a temperature driven phase transition, we expect that the complex zeros $`\beta _j,(j=1,2,\mathrm{})`$ (or at least the ones close to the real axis) condense for large enough system size on a single line. As the system size increases, those zeros will move towards the positive real $`\beta `$-axis and the corresponding value is for large system size the inverse of the physical critical temperature $`T_c`$. Crucial information on phase transitions can be obtained from the way in which the first zero approaches the real $`\beta `$-axis. However, such an analysis depends on the extrapolation towards the infinite large system and does not allow characterization of the situation in small systems such as Ala10-Gly5-Ala10. One possible extension of the above ideas to “phase transitions” in biological molecules and other small systems is the classification scheme by Borrmann et al. . In this approach one computes the discrete line density of zeros as an average of the inverse distances between neighboring zeros,

$$\varphi (\tau _k)=\frac{1}{2}\left(\frac{1}{|\beta _k\beta _{k1}|}+\frac{1}{|\beta _{k+1}\beta _k|}\right),$$
(14)
and approximates $`\varphi (\tau )`$ by a simple power law $`\varphi (\tau )\tau ^\alpha `$. Taking the first four complex zeros, one obtains

$$\alpha =\frac{\mathrm{ln}\varphi (\tau _3)\mathrm{ln}\varphi (\tau _2)}{\mathrm{ln}\tau _3\mathrm{ln}\tau _2}.$$
(15)
With a second parameter $`\gamma `$, related to the crossing angle of this line with the real axis,

$$\gamma =[\mathrm{Re}(\beta _2)\mathrm{Re}(\beta _1)]/(\tau _2\tau _1),$$
(16)
and following the classification scheme by Grossmann and Rosenhauer , phase transitions can now be classified according to the values of these two parameters: for $`\alpha 0`$ and $`\gamma =0`$ one has a phase transition of first order, it is of second order if $`0<\alpha <1`$ and arbitrary $`\gamma `$, and for $`\alpha >1`$ and arbitrary $`\gamma `$ one has a higher order transition. We have evaluated the usefulness of this approach both for spin systems and polyalanine chains . Preliminary results for Ala10-Gly5-Ala10 are also listed in Ref. .

For Ala10-Gly5-Ala10, we find two lines of complex zeros. The corresponding first four zeros for each characteristic line are listed in Table 2. Our error estimate is based on the jackknife method  with 16 bins. These lines lead to two critical temperatures $`T_{hc}=480`$ K and $`T_f=271`$ K (estimated from the real part of $`\beta `$) that agree with the corresponding values $`T_{hc}=483\pm 8`$ K and $`T_f=265\pm 7`$ K found by us above with different methods.

Using these zeros we have calculated the parameters $`\alpha `$ and $`\gamma `$ that characterize in the Borrmann et al. approach phase transitions in small systems. For the first transition, at $`T=480`$ K, we find $`\alpha =1.1(1.5)`$ and $`\gamma =0.4(2)`$. The errors reflect large fluctuations in the values of the two parameters $`\alpha `$ and $`\gamma `$ that do not allow us to determine whether the helix-coil transition is a weak first order or a strong second order phase transition. This problem was also observed in our earlier work on polyalanine  where we were also not able to establish clearly the order of the helix-coil transition. However, our results illustrate the strength of this transition that also leads to the pronounced peak in the specific heat observed in Fig. 3, and suggest a nucleation mechanism for $`\alpha `$-helix formation. Our data are more decisive in the case of the second transition, at $`T=265`$ K, which marks the collapse and folding of the peptide. Here we find $`\alpha =0.32(8)`$ and $`\gamma =0.36(2)`$. These values indicate a second-order transition which is consistent with what one would expect for a transition between extended and compact structures and imply that collapse and folding of the Ala10-Gly5-Ala10 is connected with long range correlations between the residues.

Our above analysis of the thermodynamics of our peptide suggests that Ala10-Gly5-Ala10 folds in a 2 step process. The first step is the formation of $`\alpha `$-helices and can be characterized by a helix-coil transition temperature $`T_{hc}=483\pm 8`$ K. The formation of $`\alpha `$-helices then restricts the possible configuration space. Energetically most favorable is the folding of two $`\alpha `$-helices (made out of the alanine residues) into a hairpin. This second step can be characterized by a lower folding temperature $`T_f=265\pm 7`$ K. Note that this folding temperature is in the biological relevant temperature regime while helix-formation can also happen at much higher temperatures. The above described two step folding of our artificial peptide is reminiscent of the well known framework  and collision-diffusion model  of folding which also propose that local elements of native local secondary structure form independently of tertiary structure. These elements diffuse until they collide and coalesce to give a tertiary structure. In our case, the temperature region of $`265480`$ K is the one where the thermal energy of the molecule does not allow coalescing of the helix-fragments that therefore form and decay. Some stabilization happens when these fragments try to form one extended helix, however, the inherent flexibility of the glycine residues, connecting the two alanine chains, and the gain in Lennard-Jones energy lead instead at temperatures below $`T_f`$ to a U-turn-like bundle of two (antiparallel) $`\alpha `$-helices connected by a turn of glycine residues as the most stable structure. Note that this picture is consistent with energy landscape theory and funnel concept . Fig. 5b depicts the appearance of a folding funnel at $`T=270`$ K towards our “native structure” ‘A’. The competing structure ‘B’, that at this temperature has a slightly higher ($`1k_BT`$) free energy ( see the 1 $`k_BT`$ contour line for structure ‘A’ that is missing for conformer ‘B’), acts as a local trap. However, the free energy barriers of $`6k_BT`$ can be overcome at this temperature by thermal fluctuations. Below that temperature the relative weight of structure ‘B’ decreases (see Fig.7) and its free energy difference to ‘A’ increases: the energy landscape becomes even more funnel like (data not shown). The energy landscape of Fig. 5b allows for a multitude of folding pathways that all, however, follow the above described two-step process.

An interesting question is how general the above obtained results are. A direct comparison with experimental data is difficult since solvent effects were neglected in the simulation of Ala10-Gly5-Ala10 and most experiments study solvated peptides. An exception are the techniques developed by Jarrold and collaborators for examination of gas-phase conformations of proteins and peptides . An experimental study of Ala10-Gly5-Ala10 using these techniques is now planed. The authors are not aware of experimental results for the solvated peptide. In order to compare our work with experiments of other short helical peptides , we therefore started now simulations of Ala10-Gly5-Ala10 where the solvation effects are approximated by a solvent accessible surface term . This will allows us also to test the dependency of our results on the solvation model. Simulating another, slightly more complicated, artifical peptide, Ala10-Gly5-Ala10-Gly5-Ala10 that presumably will fold in a three-helix bundle, will allow in addition a direct comparison with recent experimental work by Myers and Oas  where the relation between helix-formation and folding was studied for the 58-residue B domain of protein A.

## IV conclusion

In summary, we have performed multicanonical simulations with high statistics of a simple artificial peptide, the 25 residue Ala10-Gly5-Ala10. We found that this peptide folds into a specific structure that is determined solely by the intrinsic properties of the molecule (since solvent interactions are absent in our simulations. In gas-phase, the peptide exhibits two characteristic transitions. At $`T_{hc}=483\pm 8`$ K we observed a helix-coil transition that is either a weak first order transition or a strong second order transition. Our results indicate that there is a second transition at $`T_f=265\pm 7`$ K, the folding transition, that is second order-like. These results suggest that folding of this peptide in gas-phase is a two-step process. In a first step, the alanine residues form independently helical segments which then afterwards in a second step assemble to a U-turn like structure of two antiparallel $`\alpha `$-helices connected by a turn. By using an implicit solvent model in our simulations we started now to investigate whether the final structure or this two-step process changes in the present of water.

Acknowledgements:
U. Hansmann gratefully acknowledges support by a research grant from the National Science Foundation (CHE-9981874), and N.A. Alves support by CNPq (Brazil).

Figure Captions:

The average helicity $`<n_H>(T)`$ of Ala10-Gly5-Ala10 as a function of temperature $`T`$. The corresponding values of the susceptibility $`\chi (T)`$ are ploted in the inset. Our data rely on a single multicanonical simulation of 8,000,000 Monte Carlo sweeps.

The average ECEPP/2 energy $`<E_{tot}>`$ and the corresponding partial energy terms, Coulomb energy $`<E_C>`$, Lennard-Jones term $`<E_{LJ}>`$, hydrogen-bond energy $`<E_{hb}>`$ and torsion energy $`<E_{tor}>`$, as a function of temperature $`T`$ for Ala10-Gly5-Ala10.

The specific heat $`C(T)`$ of Ala10-Gly5-Ala10 as a function of temperature $`T`$.

The average end-to-end distance $`<d_{ee}>(T)`$ as a function of temperature $`T`$ for Ala10-Gly5-Ala10.

Free energy landscape of Ala10-Gly5-Ala10 as a function of helicity $`n_H`$ and end-to-end distance $`d_{ee}`$ at temperature (a) $`T_{hc}=480`$ K and (b) $`T_f=270`$ K. The contour lines are drawn in multiples of $`k_BT`$ indicated in the key.

Lowest-energy conformation ‘A’ of Ala10-Gly5-Ala10 (a) and the conformation ‘B’ (which has the largest helicity) (b) as found in our multicanonical simulation of 8,000,000 Monte Carlo sweeps.

Relative weight of the lowest-energy conformation ‘A’ and conformation ‘B’ (which has maximal helicity) as a function of temperature.


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# The Southern Dwarf Hunt: Local Group Dwarf Candidates in the Southern Sky1footnote 11footnote 1Based on observations made with the Isaac Newton Telescope, operated on the island of La Palma by the Isaac Newton Group in the Spanish Observatorio del Roque de los Muchachos of the Instituto de Astrofísica de Canarias, and on observations made at Cerro Tololo Interamerican Observatory (CTIO). CTIO is operated by the Association of Universities for Research in Astronomy, Inc. (AURA) under a cooperative agreement with the National Science Foundation as part of the National Optical Astronomy Observatories (NOAO).

## 1 Dwarf Galaxies

We describe here results of our search for Local Group low surface-brightness (LSB) dwarf galaxies222These three categories are formally separate. There are large LSB galaxies and bright dwarfs, and no doubt many of all sorts in the distance as well as nearby. It should be clear from the discussion below why in our work the three go together. For the sake of brevity we will generally call our targets “dwarf galaxies” and allow the “LSB” and “nearby” adjectives be implied.. LSB dwarfs form a population heavily selected against in galaxy surveys and catalogs (for obvious reasons), but of disproportionate importance in stellar, galactic and cosmological studies.

An accurate census of the small and faint end of the galaxy population is necessary for the determination of the true galaxy luminosity function, and is a difficult thing to do (McGaugh, 1996; Impey & Bothun, 1997). The Schechter form of the luminosity function with exponent $`\alpha `$ in the range 1.2-1.8, a popular region, formally diverges. If there is not to be an infinite number of faint galaxies, the function must truncate; this point has not yet been found, and faint galaxies dominate by number in volume-limited samples (Marzke & Da Costa, 1997). The luminosity function itself provides strong constraints on theories of galaxy formation (Binggeli, Sandage & Tammann, 1988). In particular, the number of dwarfs in the Local Group appears to be underpredicted by an order of magnitude in hierarchical clustering theories (Klypin et al., 1999). Although refinements in theory narrow the discrepancy (Benson et al., 2001), the problem remains (see our discussion below).

Apart from numbers, the clustering properties of dwarfs compared with larger galaxies contains important information on such things as the bias parameter and the spectrum of initial density fluctuations from which galaxies grew (Binggeli, Sandage & Tammann, 1988).

Dark matter dominates the internal dynamics of dwarfs to a much greater degree than larger objects (Mateo, 1994; Hargreaves et al., 1996), and thus they provide an especially good place to investigate the nature of this mysterious stuff. The external dynamics has been used to derive an age of the universe independent of the global Hubble constant (Lynden-Bell, 1981; Whiting, 1999) as well as the mass of the Local Group.

The star-formation histories of dwarf galaxies appear to be qualitatively different from those of larger objects; indeed, within the Local Group each dwarf differs from each other (Hodge, 1994; Grebel, 1997; Mateo, 1998). They thus provide valuable laboratories to study the general phenomenon of star formation in a variety of environments.

## 2 Dwarf Hunting

One expects LSB dwarf galaxies to be hard to find. It requires a great deal of care to detect an object which is actually fainter than the night sky (and our targets are typically only a few percent of dark sky brightness). This means that in any survey they will be seriously underrepresented. Accordingly, one must look at a lot of sky to find a few (or concentrate effort where there is likely to be a relative abundance of them). For this reason photographic plates, which provide large area coverage, have been the popular basis of searches. However, CCDs provide greater sensitivity and more controlled enhancement of small signals in large noise, so when possible they have been used. (For a discussion of selection effects affecting LSB objects, see Davies, Impey & Phillipps (1999), especially the contributions of Freeman, of McGaugh and of Huchra; also useful is the survey of surveys by Delcanton in the same volume.)

To correct for the unavoidable incompleteness and bias of a survey it is useful to have strict criteria, and if possible to perform the survey automatically. Armandroff, Davies & Jacoby (1998) and Armandroff, Jacoby & Davies (1999) have used an enhancement technique on digitized POSSII plates to discover new satellite galaxies of M31. However, any direct detection algorithm for low-signal objects tends to produce a very large number of false detections. For this reason, and to cover larger areas in the sky, visual examination of photographic sky surveys is a viable technique. Karachentseva & Karachentsev (1998), Karachentsev et al. (2000a) and Karachentseva & Karachentsev (2000) examined film copies of the POSSII plates (with some success, Karachentsev & Karachentseva (1999)), as did Côté et al. (1997).

As opposed to limited-area searches (for example, around nearby galaxy groups where the probablility of encountering a galaxy of any type is assumed to be higher), objects in the Local Group might be anywhere in the sky, requiring a full 4$`\pi `$ steradian search. In this case the requirement for coverage, together with the small physical size of CCDs, have made the use of photographic methods mandatory for our current efforts333 Great efforts are currently being made to expand the area coverage of CCD surveys. The Sloan Digital Sky Survey, to name one, covers a significant fraction of the sky. However, data reduction techniques used in (at least) the first release of Sloan data will not allow the detection of large, low surface brightness objects. All-sky searches for this type of object are several years away. We will watch with interest to see whether there is a large population of extremely low surface brightness objects visible only to deep CCD surveys.. Avoiding also the construction of an elaborate automatic algorithm, we decided to perform a visual search of survey plates starting with the southern sky.

To this end a visual examination of all 894 fields covered by the ESO-SRC and SERC Equatorial surveys of the southern sky $`(\delta <3\mathrm{°})`$ was performed. It was discovered early on that the glass copies of the surveys were significantly better than film copies for picking up very faint objects, so glass was used exclusively. Objects resembling the Andromeda dwarf spheroidals and the Tucana dwarf, that is of very low surface brightness, diffuse and large (one to a few minutes of arc), were noted. (Very nearby objects, of large angular size and completely resolved, would tend to get lost among the star foreground, so our search is not sensitive to close Milky Way satellites. We had no chance, for instance, of finding the Sagittarius dwarf visually. Paradoxically, more distant objects are easier to see.) The “bright” limit of accepted objects is difficult to quantify and probably varied significantly from plate to plate, though in all cases was well above the brightness of the known dwarfs. The faint limit, within the variations of the plate material itself, was probably more stable. (For an a posteriori examination of the limits and reliability of the survey, see section 5.1.) The size was determined with a ruler and the image on the plate, which was not always the same size as that found during followup observations and is in any case ill-defined for most of these objects. No candidate measuring one minute or over in all directions was rejected as being too small; a few cases which appeared to be on the borderline and are in fact smaller were included.

Even though LSB galaxies cannot be detected near the galactic plane, all plates were examined. At these signal levels the distribution of Galactic nebulosity dense enough to interfere with the search is very irregular and difficult to predict, so we avoided the problem of prediction altogether. As a byproduct, we discovered several planetary nebulae (which are interesting in their own right).

Plate defects were screened out by requiring an object’s presence on both blue and red plates. The same procedure eliminated most reflection nebulae due to their blue colors. Dwarflike objects noted by the UK Schmidt Telescope Unit during the survey (Tritton, 1974), but not previously followed up, were also added to the list. Catalogs of known objects were consulted (chiefly the NASA Extragalactic Database, NED) and galaxies of high radial velocity as well as planetary nebulae were excluded. Finally, when time at the telescope for followup observations permitted, we examined objects which were not necessarily LSB dwarfs but had been identified by others as possible Local Group members.

## 3 Followup Observations

The majority of the objects from our list (both those derived from survey plates and those from other sources) were imaged using the 1.5m telescope at Cerro Tololo Interamerican Observatory (CTIO) with a thinned Tek $`2048\times 2048`$ CCD. Some of the more northerly ones were followed up using the central CCD of the Wide Field Camera (WFC) on the 2.5m Isaac Newton Telescope (INT) at the Observatorio del Roque de los Muchachos in the Canary Islands. At the f/13.5 Cassegrain focus of the 1.5m CTIO telescope the scale is 0.24 arcsec/pixel and the field coverage is just over $`8\times 8`$ arcmin. The central thinned 4k$`\times `$2k EEV CCD on the INT WFC, mounted at prime focus, gives a spatial coverage of $`22\times 11`$ arcmin with a scale of 0.33 arcsec/pixel.

Candidates were initially examined by taking 15–20 minute exposures in the R band. With seeing between 0.9 - 1.4 arcsec this enables stellar objects to $`R23`$ to be detected. At this depth objects close to or within the Local Group should begin to resolve into stars, with the tip of the giant branch becoming readily visible. If a candidate appeared to resolve into stellar components, further broadband observations in V and I together with narrowband $`\mathrm{H}\alpha `$ were obtained. The raw CCD frames were processed in the standard way (bias-subtracted, trimmed and flat-fielded using twilight flats taken during the observing run) in almost real time at the telescope to aid in visual inspection of candidates. A series of standard star fields taken from Landolt (1992) were observed at intervals throughout each night.

## 4 Data on the Dwarf Candidates

Positions and characteristics of the dwarf galaxy candidates we examined are summarized in table 1. We have listed first those objects derived from our examination of survey plates, then those taken from other sources in the literature. Where the object already had a designation, we have generally refrained from giving it another; where none existed, we have constructed one based on IAU recommendations (see http://cdsweb.u-strasbg.fr/iau-spec.html). KK98 denotes objects from Karachentseva & Karachentsev (1998), KKS00 from Karachentsev et al. (2000a), and KK00 from Karachentseva & Karachentsev (2000). Where more than one designation exists for an object we have tried to list all of them (in the first column), since our experience is that different workers prefer different catalogs. The positions are good to within 0.1 minute of arc444Too much should not be made of this accuracy. For many of these objects, minutes of arc in size and very diffuse, one could reasonably choose a position for the “center” which is a large fraction of a minute away from that given here.. The size is that of the visible image at the followup telescope, not of any strictly determined isophote (and would of course be different between the CTIO 1.5m and the INT 2.5m). The nature is determined from the R-band morphology, except in the cases where time allowed an H$`\alpha `$ image to confirm (or not) emission nebulae. Additional comments on each object are presented in the next section. Images of all objects are presented in the figures. The scale varies somewhat between objects as indicated in the captions; in all, north is at the top, east to the left. In the cases of certain objects (notably the Antlia and Cetus dwarfs) CTIO images are presented even though better data are available, in the interest of providing a common comparison with the other objects.

### 4.1 Comments on Individual Objects

ESO 410G005
Although this object is clearly resolving, it is slightly too far away to be a Local Group galaxy. Karachentsev et al. (2000b) place it in the Sculptor Group on the basis of HST CMD data.

Cetus
The Cetus dwarf spheroidal is larger and much more diffuse than most of the objects in our list of candidates. For more detailed information, see Whiting, Hau & Irwin (1999).

WHI B0113-62
Nebulosity connected with the galaxy cluster Abell S0143.

WHI B0200-03
This is clearly resolving even on the survey plates. The followup images show it to be a cluster of blue stars (with a distant galaxy cluster in the background).

ESO298G033
A galaxy with a starlike nucleus and very faint disk.

PGC 009140
This is a dwarf spheroidal galaxy, beginning to resolve at the faintest levels (thus outside the Local Group, but not far).

WHI B0240-07
Possibly a LSB barred spiral. No detectable resolution.

WHI B0441+02
Irregular in shape, possibly spiral. There is some granulation which could come from HII regions.

ESO 085G088
This object is either in or behind the Large Magellanic Cloud. From its symmetrical, diffuse morphology we are inclined to call it a galaxy, rather than an LMC nebula.

WHI B0619-07
This appears to be a galaxy showing traces of resolution, though a superimposed foreground star makes study difficult.

WHI B0652+00
A faint reflection nebula.

PGC 020125
Irregular in morphology, resolving into stars clearly. However, the CMD shows visible stars to be supergiants, placing it at a distance of 2-4 Mpc. Informally designated the “Argo dwarf” (it lies in Carina, but there is already a Carina dwarf, so the disused constellation of Argo was resurrected for naming purposes).

WHI B0713-44
A round, brighter section of a more general Galactic nebulosity.

WHI B0717-07
Extremely faint round object. An image in H$`\alpha `$ shows it to be a smooth planetary.

PGC 020635
Irregular in form, with foreground stars and background galaxies making detailed study difficult.

ESO368G004
Foreground stars give an illusion of resolution at first glance.

PGC 021406
A faint, diffuse galaxy.

WHI B0740-02
The low-surface brightness object is the extended halo of a cD galaxy at the center of a cluster.

WHI B0744-05
Possibly spiral, though a foreground star makes it difficult to be sure.

WHI B0750-55
A round, diffuse galaxy.

ESO 165G006
Although cataloged as a galaxy no less than three times, this object is in fact a rather pretty planetary nebula, as shown by the hydrogen-alpha image.

WHI B0921-36
A very faint, large area of Galactic nebulosity.

ESO 126G019
This object has an overall diffuse morphology. However, at the very lowest signal level there is structure which may be the beginning of resolution.

KDG 058
Very faint, diffuse galaxy.

WHI B0959-61
A large reflection nebula. No detectable H$`\alpha `$.

Antlia
The clear resolution of Antlia into stars provides a contrast with most other objects in this catalog. Further details are provided in Whiting, Irwin & Hau (1997).

PGC 030367
Extremely large and diffuse. No detectable H$`\alpha `$.

WHI B1030-62
A relatively large emission nebula, probably a planetary, as shown by our H$`\alpha `$ image.

ESO 215G009
Diffuse galaxy.

WHI B1103-14
Across the north end of this faint, diffuse galaxy lies a chain of HII regions (confirmed by H$`\alpha `$ imaging).

WHI B1117-68
A large, dim reflection nebula.

PGC 035171
Faint, diffuse galaxy.

PGC 036594
Very faint galaxy.

WHI B1241-54
Faint reflection nebula.

WHI B1243-20
A large, faint, diffuse bit of Galactic nebulosity.

WHI B1249-33
Very large and diffuse. Noted by the UKSTU.

ESO 269G066
Diffuse galaxy.

PGC 046680
Diffuse galaxy.

PGC 048001
Galaxy behind a thick layer of foreground stars.

PGC 048178
This galaxy shows traces of a bar and ring structure.

ESO174G001
This galaxy is showing some resolution into stars at the limit of detection, but only after stacking several exposures. It is therefore too distant to be in the Local Group, but is in the Local Volume.

WHI B1414-52
A planetary nebula, well shown in our H$`\alpha `$ image.

WHI B1425-47
Faint reflection nebula.

WHI B1432-47
The object derived from the survey plate is the ill-defined central brighter region of a general nebulosity that fills the CTIO 1.5m field. There is no detectable H$`\alpha `$ emission.

WHI B1432-16
Several of the knots on the south side of this galaxy are HII regions, confirmed by an H$`\alpha `$ image.

WHI B1444-16
A bright, round object, probably a ring galaxy.

WHI B1505-67
A faint wisp, which is probably part of an even fainter complex of Galactic nebulosity.

WHI B1517-41
A very faint and ill-defined bit of Galactic nebulosity.

WHI B1603-04
Diffuse galaxy.

PGC 057387
Although catalogued as a galaxy, this is clearly a Galactic reflection nebula.

WHI B1619-67
Faint reflection nebula.

PGC 058179
This is probably an obscured galaxy, though it could possibly be a Galactic reflection nebula.

WHI B1728-08
A faint ring, shown to be a planetary by the H$`\alpha `$ image.

WHI B1751-07
Although this looks like a spiral galaxy, there is detectable H$`\alpha `$ in the same shape, so we identify it as a planetary nebula.

PGC 062147
There are possibly traces of spiral arms in this low surface brightness galaxy.

ESO 458G011
A small, heavily obscured disk galaxy.

WHI B1919-04
A heavily obscured galaxy.

WHI B1952-04
An obscured galaxy with a prominent bar and ring.

ESO 027G002
Diffuse, slightly elongated.

WHI B2212-10
A face-on disk galaxy, with very faint spiral arms.

ESO 468G020
A dwarf spheroidal galaxy, too distant to be in the Local Group but not far away. Clearly resolving. Although it lies in the general direction of the Sculptor Group, it is probably too far away to be a dynamical member.

WHI B2317-32
This is a puzzling object. On the survey plate it appears to be resolving into a handful of stars, but our image shows these to be lumps in a more general nebulosity which is of vaguely spiral form. An image in H$`\alpha `$ shows that none of these is an HII region, so it is neither a distant star-forming spiral nor a young Galactic star cluster. To add to the mystery, Côté et al. (1997) detected it in HI (their object SC 2) with a low heliocentric radial velocity of 68 km s-1. We tentatively conclude that the identification of the radial velocity with the galaxy is in error, and that this is actually a distant galaxy.

Objects from Other Sources

SC 18
Included in the Sculptor Group survey of Côté et al. (1997), though on the basis of their available data possibly nearer. Our image, although lumpy, is not resolving, so it is probably in the Sculptor Group (though not on the near side).

ESO 293G035
Identified as a dwarf galaxy in the nearby Sculptor Group by Côté et al. (1997). Our image shows a more distant object with a large, low surface brightness plume.

LGS 2
Identified by Kowal, Lo & Sargent (1978) as a possible Local Group galaxy (along with LGS 3, which is such an object). Our image shows it to be a brighter region in a large diffuse nebula, so we may answer Van den Bergh & Racine (1981) with a confident “no.”

SC 24
Also in the Sculptor Group survey of Côté et al. (1997) and thought to be possibly nearer. Our image is again lumpy, though not resolved, so it is probably also in the Sculptor Group.

ESO 352G002
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. We find a distant spiral galaxy with a superposed star (which probably provided the low radial velocity).

A0107+01
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. We find no identifiable galaxy at this position, just a star (which could have provided a low radial velocity).

ESO 416G012
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image is clearly of a background spiral, with a star superposed which probably provided the low radial velocity. A higher radial velocity has since been determined.

IC 1947
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image shows a background spiral with several neighbors. The starlike nucleus may in fact be a foreground star; this would explain the two low radial velocities in the literature. A radial velocity consistent with a much larger distance has since been determined.

ESO 056G019
Identified by Schmidt & Boller (1992) as a Local Group galaxy on the basis of radial velocity, and by NED as a part of the Large Magellanic Cloud on the same basis. Our H$`\alpha `$ image shows a very pretty emission nebula, with a bright central part about three minutes across surrounded by a faint shell about eight minutes in diameter.

ESO 318G013
Identified by Schmidt & Boller (1992) as a Local Group galaxy on the basis of radial velocity. Our image (R, V and I combined) shows a lumpy spindle, possibly beginning to resolve; however, a H$`\alpha `$ image shows that the brightest of the knots are HII regions. It is therefore well outside the Local Group. A superimposed star is probably responsible for a misleading radial velocity. (The bright H$`\alpha `$ regions should provide a good optical radial velocity for any observers who care to take up the challenge.)

ISI 1106+0257
This galaxy was chosen from Impey et al. (1996) for its low radial velocity (-69 km/s). (Note that those authors make no claim as to Local Group membership.) Our picture shows a faint barred spiral with a superimposed star; the latter is probably the source of the radial velocity.

ESO 269G070
Identified by Schmidt & Boller (1992) as a Local Group galaxy based on radial velocity. Our image shows a smooth disk galaxy with a superimposed star, which is probably the source of a misleading radial velocity (a more believable velocity has since been determined). There is a faint companion galaxy almost in contact to the west.

IC 4247
Identified by Schmidt & Boller (1992) as a Local Group galaxy based on radial velocity. Our combination of three images (total of one hour exposure) shows just the beginning of resolution. It is too far away to be in the Local Group, but is in the Local Volume.

NGC 5237
Identified by Fairall (1981) as a member of the Local Group based on radial velocity. Interestingly, identified as a member of the NGC 5128 (Centaurus) Group by Hesser et al. (1984) on the same basis. Our image shows a smooth distribution of light with no trace of resolution, which places it well beyond the Local Group.

IC 4739
Identified as a Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image clearly shows a distant spiral. A superimposed star is probably the source of an erroneous radial velocity, since corrected in the literature.

IC 4789
Identified as a Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image clearly shows a distant spiral with a superposed star, the latter probably providing a misleading radial velocity (since corrected in the literature).

IC 4937
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image clearly shows a distant edge-on spiral. A superposed star is probably responsible for a misleading radial velocity.

IC 5026
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image clearly shows a distant edge-on spiral. A superposed star is probably responsible for an anomalously low radial velocity.

A2259+12
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. We find a small, bright galaxy with no trace of resolution.

ESO 347G008
Identified as Local Group galaxy by Schmidt & Boller (1992) on the basis of radial velocity. Our image shows a very diffuse object, not resolving into stars. It is possible that very faint outlying lumps actually trace spiral arms; these (as well as two brighter objects near the center) show up as well in the H$`\alpha `$ image. If this galaxy is indeed a very low surface brightness face-on spiral, the size is actually something like 2.6 arc minutes. A higher radial velocity has since been determined.

## 5 Summary and Discussion

### 5.1 Completeness of the Local Group Census

This portion of the Dwarf Hunt was successful in its primary object, to find new Local Group dwarfs555We count Antlia as a Local Group object for convenience, even though it might not be gravitationally bound to the Group. Observationally it shares the characteristics of undisputed members, such as resolution into Giant Branch stars., as well as its secondary object, to get some idea of how many Local Group galaxies might remain to be found. Unfortunately, the completeness of the survey is difficult to quantify, for three reasons: the sensitivity of the survey plates varies with weather and the condition of the photographic emulsion; galactic extinction varies in a very complicated way with position at low signal levels; and the response of the eye to faint signals is complex and depends on many things. We consider each of these effects in turn below.

Maddox, Efstathiou, & Sutherland (1990) studied the variation in sensitivity among the 185 survey plates used for the APM Galaxy Survey. They concluded that the rms plate-to-plate variation amounted to 0.178 magnitude, and that vignetting amounted to 20% loss of light as far as 2.5o from the center of the field. Combining these we may estimate 0.3 magnitudes of variation due to plate material. Since the central (brightest) region of Cetus is 25.05 magnitudes per square arc second (and it is not the faintest object in our list) we very conservatively conclude that our survey was complete to at least 24.7 mag arcsec-2 in regions clear of Milky Way interference. In particular, we went deep enough to see the majority of very faint objects in the simulations of Benson et al. (2001).

An estimate of the fraction of dwarf galaxies hidden behind the Milky Way may be made using a grading system instituted while searching the plates. Each plate was noted as “good” (424 plates total), “troublesome” (315 plates) or “difficult” (158) based on the amount of background nebulosity interfering with detection of faint galaxies. Taking as a rough measure that no dwarfs will be seen on “difficult” plates, none over about half the area of “troublesome” plates, and that any dwarf would be seen on a “good” plate, about 35% of the sky could contain an unseen faint dwarf galaxy. With eleven of this type of galaxy known in the southern sky (Stextans A, WLM, Fornax, SagDIG, Sculptor, Sextans, Phoenix, Tucana, Carina, Antlia and Cetus), we would expect something like four more to be hiding behind the Milky Way. This estimate may be somewhat low, since nebulosity which obscures but is not luminous would not have affected the plate grading; but this is probably not a big effect. (The interesting, though not Local Group, galaxy Argo was found on a “troublesome” plate).

#### 5.1.1 Reliability of Visual Surveys

There remains the hardest quantity to estimate: the reliability of visual processing of the images. The complicated nature of visual response makes a calculation from theory impractical; the only useful estimate must come from a comparison of results of independent examinations of the same data set.

The most comparable survey to ours is that reported in Karachentseva & Karachentsev (1998), Karachentsev et al. (2000a) and Karachentseva & Karachentsev (2000). Both surveys covered the entire southern sky by visual examination of survey photographs. The object of the other group was different, however: to find dwarf galaxy candidates in the entire Local Volume, well beyond the Local Group. They thus included objects smaller than our limit and applied slightly different standards of morphology. To a first approximation, however, any object meeting our standards also met theirs. The two surveys are independent, since neither used the results of the other in compiling their lists.

Our first task is to estimate $`N`$, the total number of objects visible on the southern survey plates meeting criteria common to both searches: of very low surface brightness, one to a few minutes of arc in size, not obviously ruled out by morphology nor by high radial velocity. Of these, our survey found $`n_1`$ and the Karachentsev group $`n_2`$. The probabilies of a given object being found by each group are then $`n_1/N`$ and $`n_2/N`$, respectively. The number missed by us but found by the other group, $`m_1`$, is the probability of our missing an object, times their probability of finding it, times the number of objects:

$$m_1=\left(1\frac{n_1}{N}\right)\frac{n_2}{N}N$$
(1)
So our estimate of the total number of objects is

$$N=\frac{n_1n_2}{n_2m_1}$$
(2)
The next step is to determine these numbers.

Twenty-two objects were located by both groups. There are 29 objects found by the other group which they report as large enough (over a minute of arc in both dimensions) to fit our criteria and yet do not appear in our list. Of these, a review of Digital Sky Survey images and film copies of the Survey shows 12 too bright for us to record666We think it reasonable to assume that a galaxy with relatively high surface brightness is a big galaxy far away, based on the morphology of known members of the Local Group. “Far away” in this context may mean within the Local Volume, of course, so the Karachentsev & Karachentseva group did not apply the same restriction.. A further five were rejected by us because their known radial velocities were too high for membership in the Local Group. Six are morphologically arguable, in that they didn’t look like dwarf galaxies to us but did to the Karachentsev group. This leaves six objects meeting our selection criteria which we missed; add to these, say, half of the morphologically arguable objects, for a total of nine.

On the other hand, there are 37 objects in our survey not noted by the other group: 21 galaxies, ten reflection nebulae and six planetaries (leaving out the star cluster and the galaxy clusters). Pending a close comparison of search techniques and criteria no clear conclusions can be drawn about why these were missed. On the basis of our experience with survey films as opposed to plates, we suggest that some of the explanation might be due to the lower noise levels of the latter, allowing us to detect fainter objects. It is also probable that many, or all, of the 16 galactic objects were rejected on morphological grounds.

We use equation 2 to calculate two cases: in the first, the non-galaxy objects missing from the Karachentsev group’s listing but found by us are excluded (which would be the case, for example, if their morphological selection were perfect for these objects); in the second they are included. Thus $`n_1`$ is 43 or 59, $`n_2`$ is 31, and $`m_1`$ is 9. These result in a total number of objects of 61 and 83, respectively, of which 43 and 59 have been found777We exclude the nine objects missing from our list because (as far as we know) they havn’t yet been shown not to be Local Group dwarfs., leaving 18 or 24 objects still lurking on the plates.

Consider now that of the 43 or 59 objects examined, two have proven to be Local Group dwarfs, giving a conversion rate of .047 and .034. In each case we thus expect eight-tenths of a Local Group dwarf to remain among the objects on the plates.

Our overall estimate of Local Group dwarf galaxies remaining undiscovered in the southern sky, either hidden by the Milky Way or missed by two visual surveys, is thus about five.

Although some of the assumptions and numbers going into our numerical estimate are uncertain or arguable, the overall qualitative conclusion is very robust. Let us assume, for example, a very high conversion rate of faint objects to Local Group galaxies by including all eleven known in the south and dividing by the low estimate of 43 objects plus the nine known previously (which were not in our list). With this conversion rate of about one in five, the number of additional objects required to double the Local Group’s census, say 36, comes out to 180. For this number of objects to exist on the plates and still remain unseen our survey would have had to miss almost three times as many objects as it found, of exactly the type we were looking for. The overlap with the other group’s list would be expected to be about seven or eight, one-third of the actual overlap. And this is still not enough of an expansion of Local Group membership to agree with cosmological simulations.

Our estimate of completeness of course only takes into account those objects which fall within our search criteria. We think it quite reasonable to assume that all large, bright galaxies in the Local Group (outside the region hidden behind the Milky Way) have already been found. Our search was sensitive to dwarf galaxies as faint as any known. Nearby dwarfs could have remained hidden, if they were close and diffuse enough for their stars to be lost among the Milky Way foreground; however, the volume of space available for this is not big enough to hide many of them. Galaxies that are much more concentrated than any known, so that they fell below our size criterion; or conversely much less concentrated, so that their surface brightness fell below the plate limits, would be missed. Objects that did not have Giant Branch stars might not be seen, but that requires no star formation at all, or a strange sort which is not elsewhere seen.

### 5.2 Other Objects in the Catalog

There are a large number of Galactic reflection nebulae in our list. We attempted to avoid them by requiring any object to appear on both colors of survey plate, assuming that (blue) nebulae which are very faint on the blue plates would not appear at all on red plates. This is a reasonable assumption (Witt & Schild, 1986) and surely many were removed this way. However, apparently there are a range of colors among this kind of object and several were bright enough in the red to make it through.

Somewhat surprising are the galaxy clusters we found. These were not detected on the plate due to the concentration of galaxies, but rather due to a faint glow. Although this is generally centered on a dominant galaxy, it is very spread out and probably consists of intracluster stars not bound to any one galaxy.

As expected, several planetary nebulae were found near the Galactic plane. From their large apparent size and very low surface brightness, these must be either very old or heavily obscured (and in either case, rather close). Old planetaries are of particular interest due to their interaction with the interstellar medium (Borkowski, Sarazin & Soker, 1990), though in these cases the very low surface brightness will make further study by spectroscopy difficult.

The majority of our objects are galaxies far beyond the Local Group. These come in a variety of shapes, with diffuse dwarfs predominating but face-on disk galaxies well-represented. In many cases the nuclei of the latter are actually rather bright, but appeared starlike on the survey plates and so were not obviously extragalactic.

In addition to the list of objects derived from our own examination of survey plates, we had the opportunity to investigate several which were suggested by other workers as possible or probable Local Group galaxies. None of these turned out to be in the Local Group (though a few are probably nearby). The basis for almost all of these suggestions was radial velocity; either a low value was taken to indicate Local Group membership, or a more elaborate model (as in Schmidt & Boller (1992)) was applied. Both procedures are invalid, of course, when the velocity data point is incorrect, as it is for most of these.

This fact should not be taken as indicating a high level of error in radial velocity catalogs. The few instances highlighted here of superimposed stars giving misleading velocities are taken from many thousands of measurements, and most of these have been corrected. (Superimposed Galactic stars will, of course, only give erroneously low radial velocities to distant galaxies.)

We do wish to point out, however, the danger of forming conclusions based on a single type of data. Errors will always creep in, and if there is no independent method of detecting them they will remain. Astronomy (especially cosmology) is necessarily dependent on datasets containing sparse data on many objects and so is particular subject to this problem. We urge, whenever possible, the combination of different types of data. In what is no doubt an extreme example, by simple imaging we have reduced the population assigned by Schmidt & Boller (1992) to the Local Group by roughly 20%.

A similar problem shows up in the cases of NGC 5237 and ESO 056G019. In each, the same radial velocity was taken to indicate quite different locations by different authors. Again, imaging was able to resolve the disagreement.

### 5.3 Further Investigations

The obvious next step in the Dwarf Hunt is to extend the search to the other half of the sky. A similar visual survey of the northern sky is now being performed using POSSII plates, and we expect to report the results from it in due course. In addition, the objects from the Karachentsev & Karachentseva group which conform to our criteria but did not appear in our list should be followed up.

Meanwhile, the objects here presented offer much scope for further study. Lines of inquiry from the Galactic (interaction of old planetary nebulae with the interstellar medium) to the cosmological (formation and evolution of large, distant low surface brightness galaxies) are represented. In particular there is useful work to be done on the resolved dwarf galaxies in the Local Volume.

The authors wish to thank John Pilkington of the now defunct Royal Greenwich Observatory for digitally scanning many survey plates and Sue Tritton of the Royal Observatory Edinburgh for her patient assistance in distinguishing faint astronomical objects from plate defects. The authors are grateful for partial financial support from the Institute of Astronomy of the University of Cambridge and the Physics Department of the U. S. Naval Academy. GKTH acknowledges financial support from Chilean FONDECYT grant 1990442. In addition, we would like to express our appreciation for the dedicated help of the night assistants at Cerro Tololo. This research has made use of the NASA/IPAC Extragalactic Database (NED) which is operated by the Jet Propulsion Laboratory, California Institute of Technology, under contract with the National Aeronautics and Space Administration.