A rapid purification of alpha-actinin, filamin, and a 130,000-dalton protein from smooth muscle.
Brief, low ionic strength extraction of chicken gizzard at 37 degrees C yields a solution containing a limited number of proteins including alpha-actinin, filamin, actin, desmin, and a 130,000-dalton polypeptide. The proteins are then fractionated by Mg2+- and (NH4)2SO4-induced precipitations and by ion exchange and gel filtration column chromatography to give rise to highly purified preparations of alpha-actinin, filamin, and a 130,000-dalton protein. The alpha-actinin and filamin isolated by this scheme are "native" based upon their S20,w values and their ability to bind to F-actin. These procedures, with minor modification, can be used for the purification of alpha-actinin from skeletal muscle and non-muscle tissues as well as for the purification of filamin from non-muscle tissue.
D000185:Actinin / Q000302:isolation & purification*; D000199:Actins / Q000302:isolation & purification; D000818:Animals; D002645:Chickens; D002848:Chromatography, DEAE-Cellulose; D003285:Contractile Proteins / Q000302:isolation & purification*; D005895:Gizzard, Avian / Q000032:analysis; D008970:Molecular Weight; D009124:Muscle Proteins / Q000302:isolation & purification*; D009130:Muscle, Smooth / Q000032:analysis*; D009132:Muscles / Q000032:analysis; D011817:Rabbits
D000199:Actins; D003285:Contractile Proteins; D009124:Muscle Proteins; D000185:Actinin