A high-performance liquid chromatography assay for quantification of cardiac myosin heavy chain isoform protein expression.
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.
Lemon
Douglas D
DD
Gilead Colorado, Boulder, 80301, USA. douglas.lemon@ucdenver.edu
Papst
Philip J
PJ
Joly
Kristin
K
Plato
Craig F
CF
McKinsey
Timothy A
TA
D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods; D008297:Male; D009206:Myocardium / Q000378:metabolism; D018995:Myosin Heavy Chains / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D020033:Protein Isoforms / Q000032:analysis / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley
D020033:Protein Isoforms; D018995:Myosin Heavy Chains