Characterizing and controlling the inherent dynamics of cyclophilin-A

Protein Sci. 2009 Apr;18(4):811-24. doi: 10.1002/pro.89.

Abstract

With the recent advances in NMR relaxation techniques, protein motions on functionally important timescales can be studied at atomic resolution. Here, we have used NMR-based relaxation experiments at several temperatures and both 600 and 900 MHz to characterize the inherent dynamics of the enzyme cyclophilin-A (CypA). We have discovered multiple chemical exchange processes within the enzyme that form a "dynamic continuum" that spans 20-30 A comprising active site residues and residues proximal to the active site. By combining mutagenesis with these NMR relaxation techniques, a simple method of counting the dynamically sampled conformations has been developed. Surprisingly, a combination of point mutations has allowed for the specific regulation of many of the exchange processes that occur within CypA, suggesting that the dynamics of an enzyme may be engineered.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cyclophilin A / chemistry*
  • Cyclophilin A / genetics*
  • Humans
  • Models, Molecular
  • Nuclear Magnetic Resonance, Biomolecular
  • Point Mutation
  • Protein Conformation
  • Protein Engineering
  • Static Electricity
  • Temperature

Substances

  • Cyclophilin A