On the molecular discrimination between adenine and guanine by proteins.

The molecular recognition and discrimination of adenine and guanine ligand moieties in complexes with proteins have been studied using empirical observations on carefully selected crystal structures. The distribution of protein folds that bind these purines has been found to differ significantly from that across the whole PDB, but the most populated architectures and folds are also the most common in three genomes from the three different domains of life. The protein environments around the two nucleic acid bases were significantly different, in terms of the propensities of amino acid residues to be in the binding site, as well as their propensities to form hydrogen bonds to the bases. Plots of the distribution of protein atoms around the two purines clearly show different clustering of hydrogen bond donors and acceptors opposite complimentary acceptors and donors in the rings, with hydrophobic areas below and above the rings. However, the clustering pattern is fuzzy, reflecting the variety of ways that proteins have evolved to recognise the same molecular moiety. Furthermore, an analysis of the conservation of residues in the protein chains binding guanine shows that residues in contact with the base are in general better conserved than the rest of the chain.

Conservation helps to identify biologically relevant crystal contacts.

Some crystal contacts are biologically relevant, most are not. We assess the utility of combining measures of size and conservation to discriminate between biological and non-biological contacts. Conservation and size information is calculated for crystal contacts in 53 families of homodimers and 65 families of monomers. Biological contacts are shown to be usually conserved and typically the largest contact in the crystal. A range of neural networks accepting different combinations and encodings of this information is used to answer the following questions: (1) is a given crystal contact biological, and (2) given all crystal contacts in a homodimer, which is the biological one? Predictions for (1) are performed on both homodimer and monomer datasets. The best performing neural network combined size and conservation inputs. For the homodimers, it correctly classified 48 out of 53 biological contacts and 364 out of 366 non-biological contacts, giving a combined accuracy of 98.3 %. A more robust performance statistic, the phi-coefficient, which accounts for imbalances in the dataset, gave a value of 0.92. Taking all 535 non-biological contacts from the 65 monomers, this predictor made erroneous classifications only 4.3 % of the time. Predictions for (2) were performed on homodimers only. The best performing network achieved a prediction accuracy of 98.1 % using size information alone. We conclude that in answering question (1) size and conservation combined discriminate biological from non-biological contacts better than either measure alone. For answering question (2), we conclude that in our dataset size is so powerful a discriminant that conservation adds little predictive benefit.

Protein-protein interfaces: analysis of amino acid conservation in homodimers.

Evolutionary information derived from the large number of available protein sequences and structures could powerfully guide both analysis and prediction of protein-protein interfaces. To test the relevance of this information, we assess the conservation of residues at protein-protein interfaces compared with other residues on the protein surface. Six homodimer families are analyzed: alkaline phosphatase, enolase, glutathione S-transferase, copper-zinc superoxide dismutase, Streptomyces subtilisin inhibitor, and triose phosphate isomerase. For each family, random simulation is used to calculate the probability (P value) that the level of conservation observed at the interface occurred by chance. The results show that interface conservation is higher than expected by chance and usually statistically significant at the 5% level or better. The effect on the P values of using different definitions of the interface and of excluding active site residues is discussed.