Gilead Colorado, Boulder, 80301, USA.
Anal Biochem. 2011 Jan 1;408(1):132-5. doi: 10.1016/j.ab.2010.08.041. Epub 2010 Sep 18.
Stress signaling in the myocardium results in enhanced expression of fetal β-myosin heavy chain (β-MyHC) and reduced expression of adult α-myosin heavy chain (α-MyHC), with the net outcome of diminished myofibrillar ATPase activity and impaired contractility. Pharmacological approaches aimed at preventing this myosin isoform "switch" could provide therapeutic benefit to patients with heart failure. Myosin isoform protein expression is typically quantified using gel electrophoresis methods, which are time-consuming and prone to variability. Here we describe a facile, reversed-phase high-performance liquid chromatography (HPLC) method for rapidly determining the relative amounts of full-length α- and β-MyHC in rat hearts. The assay was validated using cardiac tissues from rats in which a key transcriptional regulator of MyHC expression, the thyroid hormone receptor, was pharmacologically manipulated. This novel assay should facilitate drug discovery efforts focused on the MyHC axis.
心肌中的应激信号导致胎β-肌球蛋白重链（β-MyHC）的表达增强，而成年α-肌球蛋白重链（α-MyHC）的表达减少，导致肌球蛋白纤维 ATP 酶活性降低和收缩功能受损。旨在预防这种肌球蛋白同工型“转换”的药物治疗方法可能会为心力衰竭患者带来治疗益处。肌球蛋白同工型蛋白的表达通常使用凝胶电泳方法进行定量，该方法耗时且容易出现变异性。在这里，我们描述了一种简便的反相高效液相色谱（HPLC）方法，可快速测定大鼠心脏中全长α-和β-MyHC 的相对含量。该测定法使用甲状腺激素受体（一种调节 MyHC 表达的关键转录调节剂）被药理学处理的大鼠心脏组织进行了验证。这种新的测定方法应该有助于专注于 MyHC 轴的药物发现工作。
