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Structure. 2017 Feb 7;25(2):276-286. doi: 10.1016/j.str.2016.12.003. Epub 2017 Jan 12.

Networks of Dynamic Allostery Regulate Enzyme Function.

Author information

1
Department of Biochemistry and Molecular Genetics, University of Colorado Denver, 12801 East 17th Avenue, MS 8101, Aurora, CO 80045, USA.
2
Department of Chemistry, Institute for Advanced Study, Technische Universität München, 85748 Garching, Germany.
3
Department of Chemistry and Biochemistry, University of Colorado, Boulder, CO 80309, USA.
4
Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, UK.
5
Department of Biochemistry and Molecular Genetics, University of Colorado Denver, 12801 East 17th Avenue, MS 8101, Aurora, CO 80045, USA. Electronic address: elan.eisenmesser@ucdenver.edu.

Abstract

Many protein systems rely on coupled dynamic networks to allosterically regulate function. However, the broad conformational space sampled by non-coherently dynamic systems has precluded detailed analysis of their communication mechanisms. Here, we have developed a methodology that combines the high sensitivity afforded by nuclear magnetic resonance relaxation techniques and single-site multiple mutations, termed RASSMM, to identify two allosterically coupled dynamic networks within the non-coherently dynamic enzyme cyclophilin A. Using this methodology, we discovered two key hotspot residues, Val6 and Val29, that communicate through these networks, the mutation of which altered active-site dynamics, modulating enzymatic turnover of multiple substrates. Finally, we utilized molecular dynamics simulations to identify the mechanism by which one of these hotspots is coupled to the larger dynamic networks. These studies confirm a link between enzyme dynamics and the catalytic cycle of cyclophilin A and demonstrate how dynamic allostery may be engineered to tune enzyme function.

KEYWORDS:

allostery; cyclophilin A; dynamics; isomerization; nuclear magnetic resonance; protein engineering

PMID:
28089447
PMCID:
PMC5336394
DOI:
10.1016/j.str.2016.12.003
[Indexed for MEDLINE]
Free PMC Article

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