title: Keep on moving: discovering and perturbing the conformational dynamics of enzymes.
abstract: CONSPECTUS: Because living organisms are in constant motion, the word "dynamics" can hold many meanings to biologists. Here we focus specifically on the conformational changes that occur in proteins and how studying these protein dynamics may provide insights into enzymatic catalysis. Advances in integrating techniques such as X-ray crystallography, nuclear magnetic resonance, and electron cryomicroscopy (cryo EM) allow us to model the dominant structures and exchange rates for many proteins and protein complexes. For proteins amenable to atomic resolution techniques, the major questions shift from simply describing the motions to discovering their role in function. Concurrently, there is an increasing need for using perturbations to test predictive models of dynamics-function relationships. Examples are the catalytic cycles of dihydrofolate reductase (DHFR) and cyclophilin A (CypA). In DHFR, mutations that alter the ability of the active site to sample productive higher energy states on the millisecond time scale reduce the rate of hydride transfer significantly. Recently identified rescue mutations restore function, but the mechanism by which they do so remains unclear. The exact role of any changes in the dynamics remains an open question. For CypA, a network of side chains that exchange between two conformations is critical for catalysis. Mutations that lock the network in one state also reduce catalytic activity. A further understanding of enzyme dynamics of well-studied enzymes such as dihydrofolate reductase and cyclophilin A will lead to improvement in ability to modulate the functions of computationally designed enzymes and large macromolecular machines. In designed enzymes, directed evolution experiments increase catalytic rates. Detailed X-ray studies suggest that these mutations likely limit the conformational space explored by residues in the active site. For proteins where atomic resolution information is currently inaccessible, other techniques such as cryo-EM and high-resolution single molecule microscopy continue to advance. Understanding the conformational dynamics of larger systems such as protein machines will likely become more accessible and provide new opportunities to rationally modulate protein function.
authors.affiliation: Department of Cellular and Molecular Pharmacology and the Howard Hughes Medical Institute, University of California , San Francisco, California 94158, United States.
authors.forename: Gira
authors.initials: G
authors.lastname: Bhabha
authors.forename: Justin T
authors.initials: JT
authors.lastname: Biel
authors.forename: James S
authors.initials: JS
authors.lastname: Fraser
mesh_terms: D021984:Cyclophilin A / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D009068:Movement; D011487:Protein Conformation; D015202:Protein Engineering; D013762:Tetrahydrofolate Dehydrogenase / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D013762:Tetrahydrofolate Dehydrogenase; D021984:Cyclophilin A