Identification of binding sites for bepridil and trifluoperazine on cardiac troponin C.

The Journal of Biological Chemistry
Q KleerekoperJ A Putkey

Abstract

The solution structure of cardiac troponin C (cTnC) (Sia, S., Li, M. X., Spyracopoulos, L., Gagne, S. M., Liu, W., Putkey, J. A. & Sykes, B. D. (1997) J. Biol. Chem. 272, 18216-18221) challenges existing structure/function models for this critical regulatory protein. For example, it is clear that the closed conformation of the regulatory N-terminal domain in Ca2+-bound cardiac troponin C (cTnC) presents a much different binding surface for Ca2+-sensitizing compounds than previously thought. We report here the use of Met methyl groups as site-specific structural markers to identify drug binding sites for trifluoperazine and bepridil on cTnC. Drug dependent changes in the NMR heteronuclear single-quantum coherence spectra of [methyl-13C]Met-labeled cTnC indicate that bepridil and trifluoperazine bind to similar sites but only in the presence of Ca2+. There are 3-4 drug binding sites in the N- and C-terminal domains of intact cTnC that exhibit fast exchange on the NMR time scale. Use of a novel spin-labeled phenothiazine and detection of isotope-filtered nuclear Overhauser effects allowed identification of drug binding sites in the shallow hydrophobic cup in the C-terminal domain and on two hydrophobic surfaces on the N-terminal r...Continue Reading

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Citations

Aug 25, 2010·Journal of Biomolecular NMR·Glenn L ButterfossRobert E London
Feb 27, 1999·Current Opinion in Biotechnology·G C Roberts
Jan 14, 2011·Biochemistry·Sandra E Pineda-SanabriaBrian D Sykes
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Apr 16, 2014·Journal of Molecular and Cellular Cardiology·Margaret V Westfall
Dec 29, 2007·Biochemical and Biophysical Research Communications·Monica X LiBrian D Sykes
Oct 11, 2016·Frontiers in Physiology·Mayra de A Marques, Guilherme A P de Oliveira
Feb 16, 2005·Journal of Muscle Research and Cell Motility·Monica X LiBrian D Sykes
Aug 14, 1999·The Journal of Biological Chemistry·Q Kleerekoper, J A Putkey
Dec 23, 2000·The Journal of Biological Chemistry·T SorsaI Kilpelainen
Feb 15, 2000·The Journal of Biological Chemistry·H PatelP D Chantler
Jun 24, 2011·Journal of Biomolecular Structure & Dynamics·Jayson F Varguhese, Yumin Li
Oct 10, 2001·FEBS Letters·E AbusamhadnehP R Rosevear
Nov 30, 2005·Biochemistry·Ryan M B HoffmanBrian D Sykes
Aug 27, 2003·Biochemistry·Douglas G WardIan P Trayer

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