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.