Evidence of chemical exchange in recombinant Major Urinary Protein and quenching thereof upon pheromone binding

J Biomol NMR. 2007 May;38(1):3-9. doi: 10.1007/s10858-006-9110-1. Epub 2007 Mar 7.

Abstract

The internal dynamics of recombinant Major Urinary Protein (rMUP) have been investigated by monitoring transverse nitrogen-15 relaxation using multiple-echo Carr-Purcell-Meiboom-Gill (CPMG) experiments. While the ligand-free protein (APO-rMUP) features extensive evidence of motions on the milliseconds time scale, the complex with 2-methoxy-3-isobutylpyrazine (HOLO-rMUP) appears to be much less mobile on this time scale. At 308 K, exchange rates k (ex) = 500-2000 s(-1) were typically observed in APO-rMUP for residues located adjacent to a beta-turn comprising residues 83-87. These residues occlude an entry to the binding pocket and have been proposed to be a portal for ligand entry in other members of the lipocalin family, such as the retinol binding protein and the human fatty-acid binding protein. Exchange rates and populations are largely uncorrelated, suggesting local 'breathing' motions rather than a concerted global conformational change.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Computer Simulation
  • Nitrogen Isotopes
  • Nuclear Magnetic Resonance, Biomolecular / methods
  • Protein Binding
  • Protein Structure, Secondary
  • Proteins / chemistry*
  • Proteins / genetics
  • Pyrazines / chemistry
  • Recombinant Proteins / chemistry*

Substances

  • Nitrogen Isotopes
  • Proteins
  • Pyrazines
  • Recombinant Proteins
  • major urinary proteins
  • 2-isobutyl-3-methoxypyrazine