High-resolution modeling of antibody structures by a combination of bioinformatics, expert knowledge, and molecular simulations.

In the second antibody modeling assessment, we used a semiautomated template-based structure modeling approach for 11 blinded antibody variable region (Fv) targets. The structural modeling method involved several steps, including template selection for framework and canonical structures of complementary determining regions (CDRs), homology modeling, energy minimization, and expert inspection. The submitted models for Fv modeling in Stage 1 had the lowest average backbone root mean square deviation (RMSD) (1.06 Å). Comparison to crystal structures showed the most accurate Fv models were generated for 4 out of 11 targets. We found that the successful modeling in Stage 1 mainly was due to expert-guided template selection for CDRs, especially for CDR-H3, based on our previously proposed empirical method (H3-rules) and the use of position specific scoring matrix-based scoring. Loop refinement using fragment assembly and multicanonical molecular dynamics (McMD) was applied to CDR-H3 loop modeling in Stage 2. Fragment assembly and McMD produced putative structural ensembles with low free energy values that were scored based on the OSCAR all-atom force field and conformation density in principal component analysis space, respectively, as well as the degree of consensus between the two sampling methods. The quality of 8 out of 10 targets improved as compared with Stage 1. For 4 out of 10 Stage-2 targets, our method generated top-scoring models with RMSD values of less than 1 Å. In this article, we discuss the strengths and weaknesses of our approach as well as possible directions for improvement to generate better predictions in the future.

Community-wide evaluation of methods for predicting the effect of mutations on protein-protein interactions.

Community-wide blind prediction experiments such as CAPRI and CASP provide an objective measure of the current state of predictive methodology. Here we describe a community-wide assessment of methods to predict the effects of mutations on protein-protein interactions. Twenty-two groups predicted the effects of comprehensive saturation mutagenesis for two designed influenza hemagglutinin binders and the results were compared with experimental yeast display enrichment data obtained using deep sequencing. The most successful methods explicitly considered the effects of mutation on monomer stability in addition to binding affinity, carried out explicit side-chain sampling and backbone relaxation, evaluated packing, electrostatic, and solvation effects, and correctly identified around a third of the beneficial mutations. Much room for improvement remains for even the best techniques, and large-scale fitness landscapes should continue to provide an excellent test bed for continued evaluation of both existing and new prediction methodologies.

Protein Loop Modeling with Optimized Backbone Potential Functions.

We represented protein backbone potential as a Fourier series. The parameters of the backbone dihedral potential were initialized to random values and optimized by Monte Carlo simulations so that generated native-like loop decoys had a lower energy than non-native decoys. The low energy regions of the optimized backbone potential were consistent with observed Ramachandran plots derived from crystal structures. The backbone potential was then used for the prediction of loop conformations (OSCAR-loop) combining with the previously described OSCAR force field, which has been shown to be very accurate in side chain modeling. As a result, the accuracy of OSCAR-loop was improved by local energy minimization based on the complete force field. The average accuracies were 0.40, 0.70, 1.10, 2.08, and 3.58 Å for 4, 6, 8, 10, and 12-residue loops, respectively, with each size being represented by 325 to 2809 targets. The accuracy was better than that of other loop modeling algorithms for short loops (<10 residues). For longer loops, the prediction accuracy was improved by concurrently sampling with a fragment-based method, Spanner. OSCAR-loop is available for download at http://sysimm.ifrec.osaka-u.ac.jp/OSCAR/ .