Chronic pressure overload induces a redistribution in myosin isoenzymes as demonstrated by Ca++-activated ATPase activity, electrophoresis under non-denaturing conditions and immunohistochemistry.
We compared, in two groups of renal hypertensive rats and control rats, the isoenzymic patterns obtained by electrophoresis under non-denaturing conditions with those observed after heavy chains digestion with S. Aureus V8 protease.
In the hypertensive animals in which a shift towards the “slow” V2 and V3 isomyosins was evident, peptide mapping always gave origin to a band which was not present in the controls.
Since we consider this peptide as a marker of the redistribution towards the “slow” isoforms, peptide mapping according to Cleveland appears to be a simple and useful method to assess differences in isomyosin composition, at least between hypertrophic pressure-overloaded and normal rat ventricles. Moreover, in our experience this technique is simple, the patterns obtained from highly purified substrates are very reproducible and the digestion allows easy and clear comparisons.
ventricular myosin pressure overload peptide mapping electrophoresis under nondenaturing conditions
This is a preview of subscription content, log in to check access.
Cleveland DW, Fischer SG, Kirschner MW, Laemmli UK (1977) Peptide mapping by limited proteolysis in sodium dodecyl sulphate and analysis by gel electrophoresis. J Biol Chem 252:1102–1106PubMedGoogle Scholar
D'Albis A, Pantaloni C, Bechet JJ (1979) An electrophoretic study of native myosin isoenzymes and of their subunit content. Eur J Biochem 99:261–266PubMedGoogle Scholar
Dalla Libera L, Sartore S, Schiaffino S (1979) Comparative analysis of chicken atrial and ventricular myosins. Biochim Biophis Acta 581:283–294Google Scholar
Flink IL, Morkin E (1977) Evidence for a new cardiac myosin species in thyrotoxic rabbit. FEBS Lett 106:197–201Google Scholar
Gorza L, Pauletto P, Pessina AC, Sartore S, Schiaffino S (1981) Isomyosin distribution in normal and pressure-overloaded rat ventricular myocardium. An immunohistochemical study. Circ Res 49:1003–1009PubMedGoogle Scholar
Hoh JFY, Mc Grath PA, Hale PT (1978) Electrophoretic analysis of multiple forms of rat cardiac myosin: effects of hypophysectomy and thyroxine replacement. J Mol Cell Cardiol 10:1053–1076PubMedGoogle Scholar
Lanzetta PA, Alvarez LJ, Reinach PS, Candia OA (1979) An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem 100:95–98PubMedGoogle Scholar
Litten RZ, Martin BJ, Low RB, Alpert NR (1982) Altered myosin isoenzyme patterns from pressure overloaded and thyrotoxic hypertrophied rabbit hearts. Circ Res 50:856–864PubMedGoogle Scholar
Lomprè AM, Schwartz K, D'Albis A, Lacombe G, van Thien N, Swynghedauw B (1979) Myosin isoenzyme redistribution in chronic heart overload. Nature 282:105–107PubMedGoogle Scholar
Mercadier JJ, Lomprè AM, Wisnewsky C, Samuel JL, Bercovici J, Swynghedauw B, Schwartz K (1981) Myosin isoenzymic changes in several models of rat cardiac hypertrophy. Circ Res 49:525–532PubMedGoogle Scholar
Pope B, Hoh JFY, Weeds A (1980) The ATPase activity of rat cardiac myosin isoenzymes. FEBS Lett 118:205–208PubMedGoogle Scholar
Rupp H (1981) The adaptive changes in the isoenzyme pattern of myosin from hypertrophied rat myocardium as a result of pressure overload and physiological training. Basic Res Cardiol 76:79–88PubMedGoogle Scholar
Rupp H (1982) Polymorphic myosin as the common determinant of myofibrillar ATPase in different haemodynamic and thyroid states. Basic Res Cardiol 77:34–46PubMedGoogle Scholar
Sartore S, Gorza L, Dalla Libera L, Schiaffino S (1979) Fractionation of rabbit ventricular myosins by affinity chromatography with insolubilized antimyosin antibodies. FEBS Lett 106:197–201PubMedGoogle Scholar
Scheuer J, Bahn AK (1979) Cardiac contractile proteins: Adenosin-triphosphatase activity and physiological function. Circ Res 45:1–12PubMedGoogle Scholar
Schwartz K, Bouveret P, Schay C, Leger J, Swynghedauw B (1978) Immunochemical evidence for species-specificity of mammalian cardiac myosin and heavy meromyosin. Biochim Biophys Acta 495:24–36Google Scholar
Schwartz K, Bouveret P, Bercovici J, Swynghedauw B (1978) Immunochemical difference between myosins from normal and hypertrophied rat hearts. FEBS Lett 93:137–140PubMedGoogle Scholar
Schwartz K, Lecarpentier Y, Martin JL, Lomprè AM, Mercadier JJ, Swynghedauw B (1981) Myosin isoenzymic distribution correlates with speed of myocardial contraction. J Mol Cell Cardiol 13:1071–1075PubMedGoogle Scholar
Schwartz K, Lomprè AM, Lacombe G, Bouveret P, Wisnewsky C, Whalen RG, D'Albis A, Swynghedauw B (1983) Cardiac myosin isoenzymic transitions in mammals. In: Alpert NR (ed) Perspectives in cardiovascular research. Vol. 7, Myocardial hypertrophy and failure. Raven Press, New York, pp 345–358Google Scholar