Satoh K, Kitahara A, Sato K
Arch Biochem Biophys. 1985 Oct;242(1):104-11. doi: 10.1016/0003-9861(85)90484-9.
Subunits of multiple molecular forms of dimeric glutathione S-transferase in rat liver cytosol were analyzed by two-dimensional gel electrophoresis (isoelectric focusing/sodium dodecyl sulfate-electrophoresis) followed by staining with Coomassie blue dye. The five subunits, Ya, Yb, Yb', Yc, and Yp (Mr's 26,500, 27,500, 27,500, 28,500, and 26,000, respectively) of seven molecular forms, A2, AC, C2, B2, BL, L2, and GST-P, were identified by comparison of molecular weights and pI values with those of purified molecular forms and by immunoadsorption of the molecular forms in the cytosol as well as those synthesized in vitro using antibodies against the seven forms. Yp is the subunit of placental glutathione S-transferase, GST-P (YpYp), which is markedly increased in carcinogen-treated rat livers [A. Kitahara et al. (1984) Cancer Res. 44, 2698-2703; K. Satoh et al. (1985) Proc. Natl. Acad. Sci. USA 82, 3964-3968]. Microheterogeneity was detectable within Yb, Yb', and Yp subunits, the different forms, termed Yb1, Yb2, Yb'1, Yb'2, and Yp1, Yp2, being similar in size but differing by approx. 0.3 pI unit within each subunit. These microheterogeneous forms were also detectable in the polypeptides translated in vitro in a rabbit reticulocyte lysate translation system from liver poly(A)-containing RNAs, suggesting that they are translatable from distinct mRNAs.
采用二维凝胶电泳（等电聚焦/十二烷基硫酸钠电泳），随后用考马斯亮蓝染料染色，分析大鼠肝细胞溶质中二聚体谷胱甘肽S-转移酶多种分子形式的亚基。通过将分子量和pI值与纯化分子形式的分子量和pI值进行比较，并利用针对七种形式的抗体对细胞溶质中以及体外合成的分子形式进行免疫吸附，鉴定出七种分子形式A2、AC、C2、B2、BL、L2和GST-P的五个亚基，即Ya、Yb、Yb'、Yc和Yp（分子量分别为26,500、27,500、27,500、28,500和26,000）。Yp是胎盘谷胱甘肽S-转移酶GST-P（YpYp）的亚基，在致癌物处理的大鼠肝脏中显著增加[A. Kitahara等人（1984年）《癌症研究》44, 2698 - 2703；K. Satoh等人（1985年）《美国国家科学院院刊》82, 3964 - 3968]。在Yb、Yb'和Yp亚基内可检测到微异质性，不同形式称为Yb1、Yb2、Yb'1、Yb'2以及Yp1、Yp2，它们大小相似，但每个亚基内pI值相差约0.3个单位。在兔网织红细胞裂解物翻译系统中，从肝脏含多聚腺苷酸的RNA体外翻译的多肽中也可检测到这些微异质形式，表明它们可从不同的mRNA翻译而来。
