Immuno-affinity purification and characterization of the alpha subunits of G0 type G proteins from various bovine tissues.
Polyclonal antibodies to the alpha subunits of G0 type G proteins (G0 alpha) were coupled to agarose gel and used to isolate G0 alpha from solubilized membranes of various bovine tissues. The cholate extract of membranes was applied to the anti-G0 alpha-agarose gel column. The column was washed extensively, then bound proteins were eluted at a neutral pH using a commercial ActiSep Elution Medium. The proteins in the eluate displayed a single band of 39 kDa on SDS-polyacrylamide gel electrophoresis. They bound to GTP gamma S and were ADP-ribosylated by pertussis toxin. The yield of the immunoreactive G0 alpha from the extract was about 40%. Isoelectric focusing, immunoassay and peptide mapping analysis of the G0 alpha-like proteins purified from the heart and adrenal medulla indicated that these proteins were very similar to the alpha subunit of a minor subtype of G0 in the brain which was previously referred to as G0 * alpha.
Asano
T
T
Department of Biochemistry, Institute for Developmental Research, Aichi Prefectural Colony.
Morishita
R
R
Kato
K
K
D000313:Adrenal Medulla / Q000737:chemistry; D000818:Animals; D000906:Antibodies; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry; D002540:Cerebral Cortex / Q000737:chemistry; D002846:Chromatography, Affinity / Q000379:methods; D003429:Cross Reactions; D019204:GTP-Binding Proteins / Q000276:immunology / Q000302:isolation & purification*; D046911:Macromolecular Substances; D008970:Molecular Weight; D009206:Myocardium / Q000737:chemistry; D010449:Peptide Mapping
D000906:Antibodies; D046911:Macromolecular Substances; D019204:GTP-Binding Proteins