Cross-linking study on protein neighborhoods at the subunit interface of rat liver ribosomes with 2-iminothiolane.
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.
D000818:Animals; D001345:Autoradiography; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D007096:Imidoesters / Q000378:metabolism*; D008099:Liver / Q000166:cytology*; D008970:Molecular Weight; D051381:Rats; D012269:Ribosomal Proteins / Q000378:metabolism*; D012270:Ribosomes / Q000378:metabolism*
D003432:Cross-Linking Reagents; D007096:Imidoesters; D012269:Ribosomal Proteins; C005905:methyl 4-mercaptobutyrimidate