Molecular mechanical differences between isoforms of contractile actin in the presence of isoforms of smooth muscle tropomyosin

Autor: Anne-Marie Lauzon, Jenna L. Blumenthal, Nedjma B. Zitouni, Apolinary Sobieszek, Genevieve Bates, Lennart Hilbert, Horia N. Roman, Michael C. Mackey
Jazyk: angličtina
Rok vydání: 2013
Předmět:
Zdroj: PLoS Computational Biology, Vol 9, Iss 10, p e1003273 (2013)
PLoS Computational Biology
ISSN: 1553-7358
Popis: The proteins involved in smooth muscle's molecular contractile mechanism – the anti-parallel motion of actin and myosin filaments driven by myosin heads interacting with actin – are found as different isoforms. While their expression levels are altered in disease states, their relevance to the mechanical interaction of myosin with actin is not sufficiently understood. Here, we analyzed in vitro actin filament propulsion by smooth muscle myosin for -actin (A), -actin-tropomyosin- (A-Tm), -actin-tropomyosin- (A-Tm), -actin (A), -actin-tropomyosin- (A-Tm), and -actin-tropomoysin- (A-Tm). Actin sliding analysis with our specifically developed video analysis software followed by statistical assessment (Bootstrapped Principal Component Analysis) indicated that the in vitro motility of A, A, and A-Tm is not distinguishable. Compared to these three ‘baseline conditions’, statistically significant differences () were: A-Tm – actin sliding velocity increased 1.12-fold, A-Tm – motile fraction decreased to 0.96-fold, stop time elevated 1.6-fold, A-Tm – run time elevated 1.7-fold. We constructed a mathematical model, simulated actin sliding data, and adjusted the kinetic parameters so as to mimic the experimentally observed differences: A-Tm – myosin binding to actin, the main, and the secondary myosin power stroke are accelerated, A-Tm – mechanical coupling between myosins is stronger, A-Tm – the secondary power stroke is decelerated and mechanical coupling between myosins is weaker. In summary, our results explain the different regulatory effects that specific combinations of actin and smooth muscle tropomyosin have on smooth muscle actin-myosin interaction kinetics.
Author Summary Dependent on the required physiological function, smooth muscle executes relatively fast contraction-relaxation cycles or maintains long-term contraction. The proteins driving contraction – amongst them actin, tropomyosin, and the contraction-driving myosin motor – can show small changes in the way they are constructed, they can be expressed as different “isoforms”. The isoforms are supposedly tailored to support the specific contraction patterns, but for tropomyosin and actin it is unclear exactly how the isoforms' differences affect the interaction of actin and myosin that generates the muscle contraction. We measured actin movement outside the cellular environment, focusing on the effects of different isoform combinations of only actin, myosin, and tropomyosin. We found that the actin isoforms cause differences in the mechanical interaction only when tropomyosin is present, not without it. Also, all different actin-tropomyosin combinations affected the mechanical interactions in a different way. In our experiments we could not directly observe the mechanical interactions of actin, tropomyosin, and myosin, so we reconstructed them in a mathematical model. With this model, we could determine in detail how the different actin-tropomyosin combinations caused the differences that we observed in our experiments.
Databáze: OpenAIRE
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