Cross-linking study on protein topography of rat liver 60 S ribosomal subunits with 2-iminothiolane.
Rat liver 60 S ribosomal subunits were modified with 2-iminothiolane. After treatment with hydrogen peroxide, the cross-linked proteins were extracted and then separated into 24 fractions by chromatography on carboxymethylcellulose. Each protein fraction was then analyzed by diagonal polyacrylamide-sodium dodecyl sulfate gel electrophoresis (Sommer, A., and Traut, R.R. (1974) Proc. Natl. Acad. Sci. U. S. A. 71, 3946-3950). The pieces of gel containing cross-linked protein spots that were shifted from the diagonal line were labeled with 125I. The labeled protein was extracted from the gel and identified by three kinds of two-dimensional gel electrophoresis, followed by autoradiography. Fifty-three cross-linked protein pairs involving 35 protein species containing two acidic proteins were identified. From these and previous results, a preliminary model of the protein topography of the 60 S ribosomal subunit was constructed and discussed in relation to other functional data on 60 S ribosomal proteins.
D000818:Animals; D002852:Chromatography, Ion Exchange; D003432:Cross-Linking Reagents / Q000494:pharmacology*; D004591:Electrophoresis, Polyacrylamide Gel; D007096:Imidoesters / Q000494:pharmacology*; D008099:Liver / Q000032:analysis*; D008958:Models, Molecular; D051381:Rats; D012269:Ribosomal Proteins / Q000032:analysis*
D003432:Cross-Linking Reagents; D007096:Imidoesters; D012269:Ribosomal Proteins; C005905:methyl 4-mercaptobutyrimidate