A high-performance liquid chromatography assay for quantification of cardiac myosin heavy chain isoform protein expression
2011
Abstract 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
myosinisoform “switch” could provide therapeutic benefit to patients with heart failure.
Myosinisoform 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.
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