Department of Molecular Physiology and Biophysics, Larner College of Medicine, University of Vermont, Burlington, VT, USA.
Cardiovascular Research Institute, University of Vermont , Burlington, VT, USA.
J Gen Physiol. 2024 Dec 2;156(12). doi: 10.1085/jgp.202413604. Epub 2024 Oct 7.
Myosin-binding protein H (MyBP-H) is a component of the vertebrate skeletal muscle sarcomere with sequence and domain homology to myosin-binding protein C (MyBP-C). Whereas skeletal muscle isoforms of MyBP-C (fMyBP-C, sMyBP-C) modulate muscle contractility via interactions with actin thin filaments and myosin motors within the muscle sarcomere "C-zone," MyBP-H has no known function. This is in part due to MyBP-H having limited expression in adult fast-twitch muscle and no known involvement in muscle disease. Quantitative proteomics reported here reveal that MyBP-H is highly expressed in prenatal rat fast-twitch muscles and larval zebrafish, suggesting a conserved role in muscle development and prompting studies to define its function. We take advantage of the genetic control of the zebrafish model and a combination of structural, functional, and biophysical techniques to interrogate the role of MyBP-H. Transgenic, FLAG-tagged MyBP-H or fMyBP-C both localize to the C-zones in larval myofibers, whereas genetic depletion of endogenous MyBP-H or fMyBP-C leads to increased accumulation of the other, suggesting competition for C-zone binding sites. Does MyBP-H modulate contractility in the C-zone? Globular domains critical to MyBP-C's modulatory functions are absent from MyBP-H, suggesting that MyBP-H may be functionally silent. However, our results suggest an active role. In vitro motility experiments indicate MyBP-H shares MyBP-C's capacity as a molecular "brake." These results provide new insights and raise questions about the role of the C-zone during muscle development.
肌球蛋白结合蛋白 H(MyBP-H)是脊椎动物骨骼肌肌节的一个组成部分,与肌球蛋白结合蛋白 C(MyBP-C)具有序列和结构域同源性。虽然肌球蛋白结合蛋白 C 的骨骼肌同工型(fMyBP-C,sMyBP-C)通过与肌节“C 带”内的肌动蛋白细丝和肌球蛋白马达相互作用来调节肌肉收缩力,但 MyBP-H 没有已知的功能。这在一定程度上是由于 MyBP-H 在成人快肌中的表达有限,并且没有已知的与肌肉疾病有关。这里报道的定量蛋白质组学显示,MyBP-H 在产前大鼠快肌和幼鱼中高度表达,这表明它在肌肉发育中具有保守作用,并促使研究人员定义其功能。我们利用斑马鱼模型的遗传控制以及结构、功能和生物物理技术的组合来研究 MyBP-H 的作用。转染的、FLAG 标记的 MyBP-H 或 fMyBP-C 都定位于幼鱼肌纤维的 C 带中,而内源性 MyBP-H 或 fMyBP-C 的遗传缺失导致另一种蛋白的积累增加,表明它们竞争 C 带结合位点。MyBP-H 是否调节 C 带的收缩性?MyBP-C 的调节功能至关重要的球状结构域缺失了 MyBP-H,这表明 MyBP-H 可能在功能上是沉默的。然而,我们的结果表明它具有活跃的作用。体外运动实验表明,MyBP-H 具有与 MyBP-C 相同的作为分子“制动器”的能力。这些结果提供了新的见解,并提出了关于 C 带在肌肉发育过程中的作用的问题。