A. F. Anglemier

Separation of Bovine Sarcoplasmic Proteins by Vertical Gel Electrophoresis

Abstract Vertical electrophoresis utilizing polyacrylamide gels varying in concentrations from 5 to 20% was employed to separate bovine sarcoplasmic proteins by both continuous and discontinuous techniques. Results of a series of electrophoretic runs indicated that a gel concentration of 10% was optimum for separating these proteins. In terms of resolution, reproducibility and sharpness of boundaries of the separated protein bands, the discontinuous technique was found to be superior to the continuous procedure. Electrophoresis of the sarcoplasmic extracts by the discontinuous technique resulted in the separation of 18 electrophoretically different sarcoplasmic proteins.

Isolation and partial characterisation of bovine rumen myosin

Myosin was isolated from rumen muscle and purified by DEAE Sephadex A-50 chromatography. The purified myosin gave only two bands on SDS gel electrophoresis, one corresponding to the heavy chain of 210 000 D and the other corresponding to a light chain of 17 000 D. The pH optimum of the rumen myosin ATPase activity was found to be at 7·6; and at pH 9·1, there was no detectable activity. The ATPase activity of the rumen myosin was found to be lower than that of the skeletal myosin. Since it is known that the light chains are located on the globular head of the myosin molecule, where the ATPase activity is found, the lower rumen myosin ATPase activity may be due to the absence of certain light chains that are commonly found in the skeletal myosin. The rumen myosin also had a lower basic amino acid content and a lower ratio of basic to acidic amino acids than the corresponding skeletal muscle counterpart.

Electrophoretic Separation of Bovine Sarcoplasmic Proteins Fractionated by DEAE-Cellulose Chromatography

Abstract Heterogeneity of bovine sarcoplasmic protein fractions obtained by DEAE-cellulose ion exchange chromatography was investigated by vertical polyacrylamide gel electrophoresis. Results of these electrophoretic analyses indicated that the five major chromatographic fractions were quite heterogeneous. Fraction areas I and III, which appeared as single chromatographic peaks, showed five and six distinct bands, respectively, when subjected to gel electrophoresis. The other chromatographic groups, fraction areas II, IV and V, were separated into five, eight and three electrophoretically different bands, respectively. Some possible reasons for the apparent chromatographic heterogeneity are discussed.