Identification of neighboring protein pairs cross-linked with dimethyl 3,3'-dithiobispropionimidate in rat liver 40S ribosomal subunits.
Rat liver 40S ribosomal subunits were treated with a bifunctional imidoester, dimethyl 3,3'-dithiobispropionimidate (DTP), and the neighboring protein pairs were identified. The cross-linked proteins were analyzed by acrylamide/SDS diagonal gel electrophoresis (Sommer & Traut (1974) Proc. Natl. Acad. Sci. U.S. 71, 3946-3950). The cross-linked components that fell off the diagonal upon adding 2-mercaptoethanol in the second dimension were labeled with 125I in the acrylamide gel and identified by two-dimensional acrylamide/urea gel electrophoresis, followed by radioautography. Considering these results and the molecular weights, we propose the following ten pairs, according to our numbering system (Terao & Ogata (1975) Biochim. Biophys. Acta 402, 219-229): S3-S5 (S3/S3a-S4), S3-S14 (S3/S3a-S14), S3-S17 (S3/S3a-S16), S5-S22 (S4-S23/S24), S10-S12 (S8-S11), S9-S16 (S9-S18), S9-S22 (S9-S23/S24), S6-S23 (S5-S25), S17-S21 (S16-S19), and S16-S26 (S18-S27). The designation according to the proposed uniform nomenclature (McConkey et al. (1979) Mol. Gen. Genet. 169, 1-6) are given in parentheses.
D000818:Animals; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D007096:Imidoesters; D007457:Iodine Radioisotopes; D007553:Isotope Labeling; D008099:Liver / Q000032:analysis*; D008970:Molecular Weight; D051381:Rats; D012269:Ribosomal Proteins / Q000032:analysis*; D012270:Ribosomes / Q000032:analysis*
D003432:Cross-Linking Reagents; D007096:Imidoesters; D007457:Iodine Radioisotopes; D012269:Ribosomal Proteins; C007471:dimethyl dithiobispropionimidate