Asano T, Morishita R, Kato K
Department of Biochemistry, Institute for Developmental Research, Aichi Prefectural Colony.
J Biochem. 1991 Oct;110(4):571-4. doi: 10.1093/oxfordjournals.jbchem.a123621.
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.
将针对G0型G蛋白（G0α）α亚基的多克隆抗体与琼脂糖凝胶偶联，用于从各种牛组织的可溶性膜中分离G0α。将膜的胆酸盐提取物应用于抗G0α-琼脂糖凝胶柱。对该柱进行广泛洗涤，然后使用商业ActiSep洗脱介质在中性pH下洗脱结合的蛋白质。洗脱液中的蛋白质在SDS-聚丙烯酰胺凝胶电泳上显示出一条39 kDa的单带。它们与GTPγS结合，并被百日咳毒素进行ADP核糖基化。提取物中免疫反应性G0α的产量约为40%。对从心脏和肾上腺髓质纯化的G0α样蛋白进行等电聚焦、免疫测定和肽图谱分析表明，这些蛋白与大脑中一种以前称为G0*α的G0次要亚型的α亚基非常相似。
