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Biochemistry (Moscow)

, Volume 73, Issue 2, pp 178–184 | Cite as

Study of the interaction between rabbit cardiac contractile and regulatory proteins. An in vitro motility assay

  • L. V. NikitinaEmail author
  • G. V. Kopylova
  • D. V. Shchepkin
  • L. B. Katsnelson
Article

Abstract

A series of experiments was performed in an in vitro motility assay with reconstructed thin filaments to obtain pCa-force relationships for cardiac isomyosins V1 and V3. Two concentrations of each isomyosin (200 and 300 μg/ml) on the surface of a flow cell were tested. Isometric force was estimated as the amount of actin-binding protein, α-actinin, stopping thin filament movement. It was found that the amount of α-actinin stopping the movement at saturating calcium concentration for V3 was twice higher than for V1 at both concentrations of isoforms. Hill coefficients of cooperativity (h) were determined for pCa-force relationships. The value of h did not differ significantly for isoforms at 300 μg/ml of protein (h was 1.56 for V1 and 1.54 for V3). However, the Hill coefficient was higher for V3 isoform at 200 μg/ml (h = 2.00 and 1.76 for V3 and V1, respectively). Importantly, the Hill coefficient increased for both isoenzymes when their concentrations were decreased. The connection between Hill coefficient and cooperative interactions between cardiac contractile and regulatory proteins is analyzed in detail.

Key words

cardiac isomyosins cooperativity between contractile and regulatory proteins 

Abbreviations

CaTnC

calcium-troponin C complex

TnC

troponin C

OQS

oxygen-quenching system

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Copyright information

© Pleiades Publishing, Ltd. 2008

Authors and Affiliations

  • L. V. Nikitina
    • 1
    Email author
  • G. V. Kopylova
    • 1
  • D. V. Shchepkin
    • 1
  • L. B. Katsnelson
    • 1
  1. 1.Institute of Immunology and PhysiologyUral Division of the Russian Academy of SciencesEkaterinburgRussia

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