Stochastic Pairwise Alignments and Scoring Methods for Comparative Protein Structure Modeling

Adam Marko, Kate Stafford, and Troy Wymore

Despite recent advances in fold recognition algorithms that identify template structures with distant homology to the target sequence, the quality of the target-template alignment can be a major problem for distantly related proteins in comparative modeling. Here we report for the first time on the use of ensembles of pairwise alignments obtained by stochastic backtracking as a means to improve three-dimensional comparative protein models. In every one of the 35 cases, the ensemble produced by the program probA resulted in alignments that were closer to the structural alignment than those obtained from the optimal alignment. In addition, we examined the lowest energy structure among these ensembles from four different structural assessment methods and compared these with the optimal and structural alignment model. The structural assessment methods consisted of the DFIRE, DOPE and ProsaII statistical potential energies, and the potential energy from the CHARMM protein force field coupled to a Generalized Born implicit solvent model. The results demonstrate that the generation of alignment ensembles through stochastic backtracking using probA combined with one of the statistical potentials for assessing three-dimensional structures can be used to improve comparative models.