Use of thin filament reconstituted muscle fibres to probe the mechanism of force generation

Masataka Kawai; Shin'ichi Ishiwata

doi:10.1007/s10974-006-9075-4

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Use of thin filament reconstituted muscle fibres to probe the mechanism of force generation

Journal article

Peer reviewed

Use of thin filament reconstituted muscle fibres to probe the mechanism of force generation

Masataka Kawai and Shin'ichi Ishiwata

Journal of muscle research and cell motility, Vol.27(5-7), pp.455-468

2006

DOI: 10.1007/s10974-006-9075-4

PMCID: PMC2896216

PMID: 16909198

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Abstract

The technique of selective removal of the thin filament by gelsolin in bovine cardiac muscle fibres, and reconstitution of the thin filament from isolated proteins is reviewed, and papers that used reconstituted preparations are discussed. By comparing the results obtained in the absence/presence of regulatory proteins tropomyosin (Tm) and troponin (Tn), it is concluded that the role of Tm and Tn in force generation is not only to expose the binding site of actin to myosin, but also to modify actin for better stereospecific and hydrophobic interaction with myosin. This conclusion is further supported by experiments that used a truncated Tm mutant and the temperature study of reconstituted fibres. The conclusion is consistent with the hypothesis that there are three states in the thin filament: blocked state, closed state, and open state. Tm is the major player to produce these effects, with Tn playing the role of Ca2+ sensing and signal transmission mechanism. Experiments that changed the number of negative charges at the N-terminal finger of actin demonstrates that this part of actin is essential to promote the strong interaction between actin and myosin molecules, in addition to the well-known weak interaction that positions the myosin head at the active site of actin prior to force generation.

Tropomyosin - physiology

Gelsolin - metabolism

Allosteric Regulation

Myocardial Contraction - physiology

Actins - metabolism

Muscle Fibers, Skeletal - metabolism

Gelsolin - pharmacology

Animals

Cattle

Myocardium - metabolism

Troponin - physiology

Cytoskeleton - metabolism

Myosins - metabolism

Binding Sites

Details

Title: Subtitle: Use of thin filament reconstituted muscle fibres to probe the mechanism of force generation
Creators: Masataka Kawai - Department of Anatomy and Cell Biology, College of Medicine, The University of Iowa, Iowa City, IA 52242, USA. masataka-kawai@uiowa.edu
Shin'ichi Ishiwata
Resource Type: Journal article
Publication Details: Journal of muscle research and cell motility, Vol.27(5-7), pp.455-468
DOI: 10.1007/s10974-006-9075-4
PMID: 16909198
PMCID: PMC2896216
NLM abbreviation: J Muscle Res Cell Motil
ISSN: 0142-4319
eISSN: 1573-2657
Publisher: Netherlands
Grant note: R01 HL070041 / NHLBI NIH HHS R01 HL070041-04 / NHLBI NIH HHS HL70041 / NHLBI NIH HHS
Language: English
Date published: 2006
Academic Unit: Anatomy and Cell Biology; Internal Medicine
Record Identifier: 9984025326602771

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