Institute of Immunology and Physiology, Russian Academy of Sciences, 620049, Yekaterinburg, Russia.
Research Center of Biotechnology, A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, 119071, Moscow, Russia.
J Muscle Res Cell Motil. 2021 Jun;42(2):343-353. doi: 10.1007/s10974-020-09593-4. Epub 2021 Jan 3.
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.
α-原肌球蛋白(Tpm1.1)的磷酸化是心肌收缩力的调节机制之一,它是心肌中主要的 Tpm 同工型之一。Tpm1.1 分子有一个磷酸化位点,即 Ser283。Tpm 的磷酸化程度随年龄的增长而降低,也会随着心脏病理的变化而变化。心肌病理,特别是缺血,通常伴随着心肌细胞胞质 pH 值降低。我们研究了酸中毒对 S283D 取代的假磷酸化 Tpm1.1 形式的结构和功能特性的影响。我们发现,在酸中毒条件下,S283D Tpm 分子的 N-端和 C-端之间的相互作用降低,而 WT Tpm 则没有变化。pH 值降低会更大程度地提高 F-肌动蛋白与 S283D Tpm 形成的复合物的热稳定性,而与 WT Tpm 相比则没有变化。我们使用 NEM 修饰的肌球蛋白作为负载的体外运动分析,评估了 Tpm 假磷酸化对肌球蛋白与肌动蛋白相互作用产生力的影响。在酸中毒条件下,与含有 S283D Tpm 的薄丝相互作用的肌球蛋白产生的力比与含有 WT Tpm 的薄丝相互作用的力更高。此外,假磷酸化增加了肌球蛋白抵抗负载的能力。我们得出结论,缺血改变了磷酸化 Tpm 对心肌收缩功能的影响。