Partial Order Optimum Likelihood (POOL): Maximum Likelihood Prediction of Protein Active Site Residues Using 3D Structure and Sequence Properties

A new monotonicity-constrained maximum likelihood approach, called Partial Order Optimum Likelihood (POOL), is presented and applied to the problem of functional site prediction in protein 3D structures, an important current challenge in genomics. The input consists of electrostatic and geometric properties derived from the 3D structure of the query protein alone. Sequence-based conservation information, where available, may also be incorporated. Electrostatics features from THEMATICS are combined with multidimensional isotonic regression to form maximum likelihood estimates of probabilities that specific residues belong to an active site. This allows likelihood ranking of all ionizable residues in a given protein based on THEMATICS features. The corresponding ROC curves and statistical significance tests demonstrate that this method outperforms prior THEMATICS-based methods, which in turn have been shown previously to outperform other 3D-structure-based methods for identifying active site residues. Then it is shown that the addition of one simple geometric property, the size rank of the cleft in which a given residue is contained, yields improved performance. Extension of the method to include predictions of non-ionizable residues is achieved through the introduction of environment variables. This extension results in even better performance than THEMATICS alone and constitutes to date the best functional site predictor based on 3D structure only, achieving nearly the same level of performance as methods that use both 3D structure and sequence alignment data. Finally, the method also easily incorporates such sequence alignment data, and when this information is included, the resulting method is shown to outperform the best current methods using any combination of sequence alignments and 3D structures. Included is an analysis demonstrating that when THEMATICS features, cleft size rank, and alignment-based conservation scores are used individually or in combination THEMATICS features represent the single most important component of such classifiers.Author Summary: Genome sequencing has revealed the codes for thousands of previously unknown proteins for humans and for hundreds of other species. Many of these proteins are of unknown or unclear function. The information contained in the genome sequences holds tremendous potential benefit to humankind, including new approaches to the diagnosis and treatment of disease. In order to realize these benefits, a key step is to understand the functions of the proteins for which these genes hold the code. A first step in understanding the function of a protein is to identify the functional site, the local area on the surface of a protein where it affects its functional activity. This paper reports on a new computational methodology to predict protein functional sites from protein 3D structures. A new machine learning approach called Partial Order Optimum Likelihood (POOL) is introduced here. It is shown that POOL outperforms previous methods for the prediction of protein functional sites from 3D structures.