Inference of Functionally-Relevant N-acetyltransferase Residues Based on Statistical Correlations

Over evolutionary time, members of a superfamily of homologous proteins sharing a common structural core diverge into subgroups filling various functional niches. At the sequence level, such divergence appears as correlations that arise from residue patterns distinct to each subgroup. Such a superfamily may be viewed as a population of sequences corresponding to a complex, high-dimensional probability distribution. Here we model this distribution as hierarchical interrelated hidden Markov models (hiHMMs), which describe these sequence correlations implicitly. By characterizing such correlations one may hope to obtain information regarding functionally-relevant properties that have thus far evaded detection. To do so, we infer a hiHMM distribution from sequence data using Bayes’ theorem and Markov chain Monte Carlo (MCMC) sampling, which is widely recognized as the most effective approach for characterizing a complex, high dimensional distribution. Other routines then map correlated residue patterns to available structures with a view to hypothesis generation. When applied to N-acetyltransferases, this reveals sequence and structural features indicative of functionally important, yet generally unknown biochemical properties. Even for sets of proteins for which nothing is known beyond unannotated sequences and structures, this can lead to helpful insights. We describe, for example, a putative coenzyme-A-induced-fit substrate binding mechanism mediated by arginine residue switching between salt bridge and π-π stacking interactions. A suite of programs implementing this approach is available (psed.igs.umaryland.edu).Author Summary: Protein sequence data, when gathered in great quantity, contain important but implicit biological information manifest as statistical correlations. Here we describe an approach to access this information by comprehensively modeling and characterizing the distribution of sequences belonging to a major protein superfamily. This approach takes as input a large set of unaligned sequences belonging to the superfamily. By applying the minimum description length principle, it seeks the statistical model that best explains the sequences while avoiding over-fitting the data. It concurrently aligns the sequences and, to model evolutionary divergence, partitions them into subgroups that are hierarchically-arranged based upon correlated residue patterns. Auxiliary routines create PyMOL scripts to visualize the locations of correlated residues within available structures. Because these correlations likely arise from structural and biochemical constraints, they can help elucidate protein properties important for functional specificity. Comparing and contrasting sequence and structural features in this way may therefore suggest, in the light of published studies, plausible biological hypotheses for experimental investigation. We illustrate this approach with N-acetyltransferases.