An electrophoretic procedure for detecting proteins that bind actin monomers.
The electrophoretic mobility of fluorescently labeled G-actin in polyacrylamide gels under nondenaturing conditions is altered by the formation of complexes with actin-binding proteins. This effect offers a convenient method for detecting and quantitating such proteins in tissue fractions and for monitoring their purification. When followed by second-dimension electrophoresis in the presence of sodium dodecyl sulfate, the method also gives the apparent molecular weights of the actin-binding components and the stoichiometry of the complexes. The method has also been used to identify actin-binding fragments in digests of actin-binding proteins, to investigate the formation of multicomponent complexes, and to determine the calcium-sensitivity of complexes.
Safer
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Department of Anatomy, School of Medicine, University of Pennsylvania, Philadelphia 19104-6058.
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002135:Calcium-Binding Proteins / Q000032:analysis; D002645:Chickens; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005456:Fluorescent Dyes; D018260:Gelsolin; D008840:Microfilament Proteins / Q000032:analysis / Q000378:metabolism*; D009124:Muscle Proteins / Q000032:analysis; D014809:Vitamin D-Binding Protein / Q000032:analysis
D000199:Actins; D002135:Calcium-Binding Proteins; D005456:Fluorescent Dyes; D018260:Gelsolin; D008840:Microfilament Proteins; D009124:Muscle Proteins; D014809:Vitamin D-Binding Protein