Uchiumi T, Kikuchi M, Ogata K
J Biol Chem. 1986 Jul 25;261(21):9663-7.
Rat liver 80 S ribosomes were cross-linked with 2-iminothiolane. Proteins extracted from the cross-linked 80 S ribosomes were separated into 25 fractions by chromatography on carboxy methylcellulose. Each protein fraction was analyzed by diagonal polyacrylamide-sodium dodecyl sulfate gel electrophoresis. Eight pairs characteristic of 80 S ribosomes were detected which did not appear when isolated 40 S and 60 S subunits were cross-linked, and the cross-linked proteins were analyzed in similar manners. The cross-linked components were radioiodinated and then analyzed by two-dimensional gel electrophoresis, followed by autoradiography. Eight kinds of cross-links between 60 S subunit proteins and 40 S subunit proteins were identified as follows: SA30 (acidic protein with Mr 30,000)-LA33 (acidic protein with Mr 33,000), S2-LA33, S2-L11, S3a-L11, S4-L5, S25-L5, S4-L24 and S6-L24.
用2-亚氨基硫杂环戊烷对大鼠肝脏80S核糖体进行交联。从交联的80S核糖体中提取的蛋白质通过羧甲基纤维素柱色谱分离成25个组分。每个蛋白质组分通过对角线聚丙烯酰胺-十二烷基硫酸钠凝胶电泳进行分析。检测到8对80S核糖体特有的交联对，而分离的40S和60S亚基交联时未出现这些交联对，并且以类似方式分析交联蛋白。将交联组分进行放射性碘化，然后通过二维凝胶电泳分析，随后进行放射自显影。鉴定出60S亚基蛋白与40S亚基蛋白之间的8种交联如下：SA30（分子量为30,000的酸性蛋白）-LA33（分子量为33,000的酸性蛋白）、S2-LA33、S2-L11、S3a-L11、S4-L5、S25-L5、S4-L24和S6-L24。
