Isolation and characterization of one isoform of actin from cultured soybean cells.
Cultured soybean cells (SB-1 cell line) were plasmolyzed and lyophilized. Extraction of the dried powder and fractionation yielded a polypeptide with the following key properties: (a) it has a molecular weight of approximately 45,000 and an isoelectric point of approximately 5.9; (b) it is immunologically cross-reactive with rabbit antibodies affinity purified against the Mr 45,000 polypeptide of calf thymus actin; (c) it is eluted from a DEAE-cellulose column at the same ionic strength as Acanthamoeba actin; (d) it yields peptide maps, after limited proteolysis with V8 protease, similar if not identical to those of rabbit muscle actin; and (e) it binds specifically to deoxyribonuclease I. These molecular and binding properties indicate that we have purified one isoform of actin from soybean cells.
Villanueva
M A
MA
Department of Biochemistry, Michigan State University, East Lansing 48824.
Ho
S C
SC
Wang
J L
JL
D000199:Actins / Q000276:immunology / Q000302:isolation & purification*; D002460:Cell Line; D002846:Chromatography, Affinity; D002848:Chromatography, DEAE-Cellulose; D003850:Deoxyribonuclease I; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007525:Isoelectric Focusing; D008970:Molecular Weight; D010449:Peptide Mapping; D013025:Glycine max / Q000378:metabolism*
D000199:Actins; D003850:Deoxyribonuclease I