Spectroscopic and ITC study of the conformational change upon Ca2+-binding in TnC C-lobe and TnI peptide complex from Akazara scallop striated muscle

Biochem Biophys Res Commun. 2008 Apr 25;369(1):109-14. doi: 10.1016/j.bbrc.2007.11.124. Epub 2007 Dec 3.

Abstract

Akazara scallop (Chlamys nipponensis akazara) troponin C (TnC) of striated adductor muscle binds only one Ca(2+) ion at the C-terminal EF-hand motif (Site IV), but it works as the Ca(2+)-dependent regulator in adductor muscle contraction. In addition, the scallop troponin (Tn) has been thought to regulate muscle contraction via activating mechanisms that involve the region spanning from the TnC C-lobe (C-lobe) binding site to the inhibitory region of the TnI, and no alternative binding of the TnI C-terminal region to TnC because of no similarity between second TnC-binding regions of vertebrate and the scallop TnIs. To clarify the Ca(2+)-regulatory mechanism of muscle contraction by scallop Tn, we have analyzed the Ca(2+)-binding properties of the complex of TnC C-lobe and TnI peptide, and their interaction using isothermal titration microcalorimetry, nuclear magnetic resonance, circular dichroism, and gel filtration chromatography. The results showed that single Ca(2+)-binding to the Site IV leads to a structural transition not only in Site IV but also Site III through the structural network in the C-lobe of scallop TnC. We therefore assumed that the effect of Ca(2+)-binding must lead to a change in the interaction mode between the C-lobe of TnC and the TnI peptide. The change should be the first event of the transmission of Ca(2+) signal to TnI in Tn ternary complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Binding Sites
  • Calcium / chemistry*
  • Calcium / metabolism*
  • Cells, Cultured
  • Magnetic Resonance Spectroscopy
  • Muscle, Skeletal / physiology*
  • Muscle, Skeletal / ultrastructure*
  • Pectinidae
  • Protein Binding
  • Protein Conformation
  • Troponin C / metabolism*
  • Troponin C / ultrastructure*
  • Troponin I / metabolism*
  • Troponin I / ultrastructure*

Substances

  • Troponin C
  • Troponin I
  • Calcium