Immuno-Affinity Purification and Characterization of the α Subunits of Go Type G Proteins from Various Bovine Tissues
1991 年 110 巻 4 号 p. 571-574
詳細
1991 年 110 巻 4 号 p. 571-574
Polyclonal antibodies to the α subunits of Go type G proteins (Goα) were coupled to agarose gel and used to isolate Goα from solubilized membranes of various bovine tissues. The cholate extract of membranes was applied to the anti-Goα-agarose gel column. The column was washed extensively, then bound proteins were eluted at a neutral pH using a commer-cial ActiSep Elution Medium. The proteins in the eluate displayed a single band of 39 kDa on SDS-polyacrylamide gel electrophoresis. They bound to GTPγS and were ADP-ribosylated by pertussis toxin. The yield of the immunoreactive Goα from the extract was about 40%. Isoelectric focusing, immunoassay and peptide mapping analysis of the Goα-like proteins purified from the heart and adrenal medulla indicated that these proteins were very similar to the α subunit of a minor subtype of Go in the brain which was previously referred to as Go*α
