Bulk isolation of the 20,000-Da light chain of smooth muscle myosin: separation of the unphosphorylated and phosphorylated species.
The procedure of W. T. Perrie and S. V. Perry (1970, Biochem. J. 119, 31-38) has been improved and extended to allow a convenient large-scale isolation of the 20,000-Da light chain of vertebrate smooth muscle myosin. The method utilizes as source material tropomyosin-free actomyosin or myosin. The relatively pure light chain isolated from this material could be obtained in pure form by a single gel-filtration step. Separation of the unphosphorylated and phosphorylated light chain species was achieved by subsequent chromatography on a DEAE column. The solubility properties of this light chain, relevant to its use in myosin light chain kinase assays, were also established.
Sobieszek
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Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg.
D000818:Animals; D002645:Chickens; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003956:Dialysis; D004586:Electrophoresis / Q000379:methods; D009130:Muscle, Smooth / Q000032:analysis* / Q000201:enzymology; D009218:Myosins / Q000302:isolation & purification*; D010750:Phosphoproteins / Q000302:isolation & purification; D013552:Swine; D014422:Turkeys
D010750:Phosphoproteins; D009218:Myosins