Myosin binding-induced cooperative activation of the thin filament in cardiac myocytes and skeletal muscle fibers

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Abstract

Myosin binding-induced activation of the thin filament was examined in isolated cardiac myocytes and single slow and fast skeletal muscle fibers. The number of cross-bridge attachments was increased by stepwise lowering of the [MgATP] in the Ca(2+)-free solution bathing the preparations. The extent of thin filament activation was determined by monitoring steadystate isometric tension at each MgATP concentration. As pMgATP (where pMgATP is -log [MgATP]) was increased from 3.0 to 8.0, isometric tension increased to a peak value in the pMgATP range of 5.0–5.4. The steepness of the tension-pMgATP relationship, between the region of the curve where tension was zero and the peak tension, is hypothesized to be due to myosin-induced cooperative activation of the thin filament. Results showed that the steepness of the tension-pMgATP relationship was markedly greater in cardiac as compared with either slow or fast skeletal muscle fibers. The steeper slope in cardiac myocytes provides evidence of greater myosin binding-induced cooperative activation of the thin filament in cardiac as compared with skeletal muscle, at least under these experimental conditions of nominal free Ca2+. Cooperative activation is also evident in the tension-pCa relation, and is dependent upon thin filament molecular interactions, which require the presence of troponin C. Thus, it was determined whether myosin-based cooperative activation of the thin filament also requires troponin C. Partial extraction of troponin C reduced the steepness of the tension-pMgATP relationship, with the effect being significantly greater in cardiac than in skeletal muscle. After partial extraction of troponin C, muscle type differences in the steepness of the tension-pMgATP relationship were no longer apparent, and reconstitution with purified troponin C restored the muscle lineage differences. These results suggest that, in the absence of Ca2+, myosin-mediated activation of the thin filament is greater in cardiac than in skeletal muscle, and this apparent cooperativity requires the presence of troponin C on thin filament regulatory strands.

Original languageEnglish (US)
Pages (from-to)1430-1442
Number of pages13
JournalBiophysical journal
Volume68
Issue number4
DOIs
StatePublished - 1995

Bibliographical note

Funding Information:
I thank Drs. Richard Moss, John Faulkner, and Margaret Westfall for helpful comments on earlier versions of this manuscript. I also thank Drs. Richard Moss and Marion Greaser for their kind gift of purified troponin C. This work was supported in part by grants from National Institutes ofHealth, Whitaker Foundation, American Heart Association of Michigan, University of Michigan Office of Vice President for Research, and by a Rackham Faculty grant. J. M. Metzger is an Established Investigator of the American Heart Association.

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