Coupling Protein Side-Chain and Backbone Flexibility Improves the Re-design of Protein-Ligand Specificity.
Coupling Protein Side-Chain and Backbone Flexibility Improves the Re-design of Protein-Ligand Specificity.
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DOI：
10.1371/journal.pcbi.1004335
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发表时间：
2015
影响因子：
4.3
通讯作者：
Kortemme T
中科院分区：
文献类型：
--
作者：
Ollikainen N;de Jong RM;Kortemme T
Interactions between small molecules and proteins play critical roles in regulating and facilitating diverse biological functions, yet our ability to accurately re-engineer the specificity of these interactions using computational approaches has been limited. One main difficulty, in addition to inaccuracies in energy functions, is the exquisite sensitivity of protein–ligand interactions to subtle conformational changes, coupled with the computational problem of sampling the large conformational search space of degrees of freedom of ligands, amino acid side chains, and the protein backbone. Here, we describe two benchmarks for evaluating the accuracy of computational approaches for re-engineering protein-ligand interactions: (i) prediction of enzyme specificity altering mutations and (ii) prediction of sequence tolerance in ligand binding sites. After finding that current state-of-the-art “fixed backbone” design methods perform poorly on these tests, we develop a new “coupled moves” design method in the program Rosetta that couples changes to protein sequence with alterations in both protein side-chain and protein backbone conformations, and allows for changes in ligand rigid-body and torsion degrees of freedom. We show significantly increased accuracy in both predicting ligand specificity altering mutations and binding site sequences. These methodological improvements should be useful for many applications of protein – ligand design. The approach also provides insights into the role of subtle conformational adjustments that enable functional changes not only in engineering applications but also in natural protein evolution. Designing new protein–ligand interactions has tremendous potential for engineering sensitive biosensors for diagnostics or new enzymes useful in biotechnology, but these applications are extremely challenging, both because of inaccuracies of the energy functions used in modeling and design, and because protein active and binding sites are highly sensitive to subtle changes in structure. Here we describe a new method that addresses the second problem and couples changes in the structure of the protein backbone and of the amino acid side chains, the amino acid sequence, and the conformation of the ligand and its orientation in the binding site. We show that our method improvements significantly increase the accuracy of designing protein–ligand interactions compared to current state-of-the-art design methods. We assess these improvements in two important tests: the first predicts mutations that change ligand-binding preferences in enzymes, and the second predicts protein sequences that bind a given ligand. In these tests, subtle conformational changes made in our model are essential to recapitulate both the results from engineering experiments and the sequence diversity occurring in natural protein families. These results therefore shed light on the mechanisms of how new protein functions might have emerged and can be engineered in the laboratory.
