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Conformation of a Troponin-I Peptide Bound to Troponin-C as Determined by 1H NMR

  • A. Patricia Campbell
  • Brian D. Sykes
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 255)

Abstract

The Ca+2-mediated interaction between troponin-l (Tn-I) and troponin-C (Tn-C) in the thin filament of skeletal muscle is an important element in the regulation of the actomyosin ATPase. It has been proposed that the structural changes produced by Ca+2 binding result in the presentation of regions on the surface of Tn-C capable of binding Tn-I with high affinity (1,2,3). In particular, it appears that residues 84–1151 on Tn-C (which represent the N-terminal side of Ca+2-binding site III) interact with residues 97–117 of Tn-I (3,5). A synthetic analog of the inhibitory region of Tn-I, Nα-acetyl-Tn-l(104–115)amide, has been made by solid phase synthesis. This region represents the minimum sequence necessary for inhibition of actomyosin ATPase activity and is found to bind to Tn-C in a Ca+2-dependent fashion, with a binding constant of 1 x 105M-1 (6,7). The following body of work primarily addresses the question of secondary structure adopted by the Tn-I peptide when bound to Tn-C. In order to probe the structure of the bound Tn-I peptide, high field two-dimensional Nuclear Magnetic Resonance (NMR) techniques are used. Possible binding sites on the Tn-C molecule are also considered in light of recent findings from cross-linking studies between rabbit s-Tn-C, to which a maleimide cross-linker molecule has been attached at Cys98, and residues on whole Tn-I (8).

Keywords

Nuclear MagnetiC Resonance Distance Restraint Actomyosin ATPase Interproton Distance Ring Pucker 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

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Copyright information

© Plenum Press, New York 1989

Authors and Affiliations

  • A. Patricia Campbell
    • 1
  • Brian D. Sykes
    • 1
  1. 1.Department of Biochemistry, and MRC Group in Protein Structure and FunctionUniversity of AlbertaEdmontonCanada

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