Calcium-induced flexibility changes in the troponin C-troponin I complex

Xinmei Zhao, Tomoyoshi Kobayashi, Zygmunt Gryczynski, Ignacy Gryczynski, Joseph Lakowicz, Robert Wade, John H. Collins

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

The contraction of vertebrate striated muscle is modulated by Ca2+ binding to the regulatory protein troponin C (TnC). Ca2+ binding causes conformational changes in TnC which alter its interaction with the inhibitory protein troponin I (TnI), initiating the regulatory process. We have used the frequency domain method of fluorescence resonance energy transfer (FRET) to measure distances and distance distributions between specific sites in the TnC-TnI complex in the presence and absence of Ca2+ or Mg2+. Using sequences based on rabbit skeletal muscle proteins, we prepared functional, binary complexes of wild-type TnC and a TnI mutant which contains no Cys residues and a single Trp residue at position 106 within the TnI inhibitory region. We used TnI Trp-106 as the FRET donor, and we introduced energy acceptor groups into TnC by labeling at Met-25 with dansyl aziridine or at Cys-98 with N-(iodoacetyl)-N'-(1-sulfo-5-naphthyl)ethylenediamine. Our distance distribution measurements indicate that the TnC-TnI complex is relatively rigid in the absence of Ca2+, but becomes much more flexible when Ca2+ binds to regulatory sites in TnC. This increased flexibility may be propagated to the whole thin filament, helping to release the inhibition of actomyosin ATPase activity and allowing the muscle to contract. This is the first report of distance distributions between TnC and TnI in their binary complex. Copyright (C) 2000 Elsevier Science B.V.

Original languageEnglish
Pages (from-to)247-254
Number of pages8
JournalBiochimica et Biophysica Acta - Protein Structure and Molecular Enzymology
Volume1479
Issue number1-2
DOIs
StatePublished - 15 Jun 2000

Keywords

  • Calcium
  • Distance distribution
  • Fluorescence
  • Muscle
  • Regulation
  • Troponin

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