Quantitative determination of conformational, dynamic, and kinetic parameters of a ligand-protein/DNA complex from a complete relaxation and conformational exchange matrix analysis of intermolecular transferred NOESY

Hunter N.B. Moseley, Weontae Lee, Cheryl H. Arrowsmith, N. Rama Krishna

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

We report a quantitative analysis of the 13C-edited intermolecular transferred NOESY (inter-TrNOESY) spectrum of the trp-repressor/operator complex (trp-rep/op) with [ul-13C/15N]-L-tryptophan corepressor using a computer program implementing complete relaxation and conformational exchange matrix (CORCEMA) methodology [Moseley et al. (1995) J. Magn. Resort. 108B, 243-261]. Using complete mixing time curves of three inter-TrNOESY peaks between the tryptophan and the Trp-rep/op, this self-consistent analysis determined the correlation time of the bound species (τ(B) = 13.5 ns) and the exchange off-rate (k(off) = 3.6 s-1) of the corepressor. In addition, the analysis estimated the correlation time of the free species (τ(F) ~ 0.15 ns). Also, we demonstrate the sensitivity of these inter-TrNOESY peaks to several factors including the k(off) and orientation of the tryptophan corepressor within the binding site. The analysis indicates that the crystal structure orientation for the corepressor is compatible with the solution NMR data.

Original languageEnglish
Pages (from-to)5293-5299
Number of pages7
JournalBiochemistry
Volume36
Issue number18
DOIs
Publication statusPublished - 1997 May 6

All Science Journal Classification (ASJC) codes

  • Biochemistry

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