Abstract
Phosphorylation of α-tropomyosin (Tpm1.1), a predominant Tpm isoform in the myocardium, is one of the regulatory mechanisms of the heart contractility. The Tpm 1.1 molecule has one site of phosphorylation, Ser283. The degree of the Tpm phosphorylation decreases with age and also changes in heart pathologies. Myocardial pathologies, in particular ischemia, are usually accompanied by pH lowering in the cardiomyocyte cytosol. We studied the effects of acidosis on the structural and functional properties of the pseudo-phosphorylated form of Tpm1.1 with the S283D substitution. We found that in acidosis, the interaction of the N- and C-ends of the S283D Tpm molecules decreases, whereas that of WT Tpm does not change. The pH lowering increased thermostability of the complex of F-actin with S283D Tpm to a greater extent than with WT Tpm. Using an in vitro motility assay with NEM- modified myosin as a load, we assessed the effect of the Tpm pseudo-phosphorylation on the force of the actin-myosin interaction. In acidosis, the force generated by myosin in the interaction with thin filaments containing S283D Tpm was higher than with those containing WT Tpm. Also, the pseudo-phosphorylation increased the myosin ability to resist a load. We conclude that ischemia changes the effect of the phosphorylated Tpm on the contractile function of the myocardium.
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Acknowledgements
The authors thank Dr. Vera Borzova for assistance in the viscosity measurements and A. Kochurova for assistance in an in vitro motility assay experiments.
Funding
This work was funded by the Russian Foundation for Basic Research Grants 17-00-00065 (D.L.), 17-00-00070 (S.B.), 20-04-00130 (S.B.), and 18-34-20085 (D.S.); and State Program AAAA-A19-119010590010-3 (D.L.), and AAAA-A18-118020590135-3 (S.B.). This work was performed using the equipment of the Shared Research Center of Scientific Equipment of IIP UrB RAS. DSC measurements were carried out on the equipment of the Shared-Access Equipment Centre «Industrial Biotechnology» of Federal Research Center «Fundamentals of Biotechnology» of the Russian Academy of Sciences.
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Kopylova, G.V., Matyushenko, A.M., Berg, V.Y. et al. Acidosis modifies effects of phosphorylated tropomyosin on the actin-myosin interaction in the myocardium. J Muscle Res Cell Motil 42, 343–353 (2021). https://doi.org/10.1007/s10974-020-09593-4
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DOI: https://doi.org/10.1007/s10974-020-09593-4