Open AccessThe Use of a Cleavable Crosslinking Reagent to Identify Neighboring Proteins in the 30‐S Ribosomal Subunit of Escherichia coli
Author(s) -
PERETZ Hava,
TOWBIN Harry,
ELSON David
Publication year - 1976
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1976.tb10210.x
Subject(s) - ribosome , escherichia coli , reagent , protein subunit , ribosomal protein , ribosomal rna , chemistry , polyacrylamide gel electrophoresis , escherichia , biochemistry , microbiology and biotechnology , biology , enzyme , rna , gene , organic chemistry
A cleavable bifunctional reagent, dimethyl 3,3′‐dithiobispropionimidate, has been used to crosslink proteins that occupy neighboring positions in the 30‐S ribosomal subunit of Escherichia coli . The crosslinked proteins were identified, fully or partly, by their positions in two twodimensional gel electrophoretic systems, one diagonal and the other quasi‐diagonal, in which the complexes were cleaved alter the first‐dimensional run. It was found to be necessary to block the protein sulfhydryl groups in order to prevent artifactual disulfide crosslinking after extraction of the protein from the ribosome. Eleven crosslinked complexes were detected. Four were fully identified: the triplet S4‐S5‐S8, and the pairs S2‐S3, S4‐S5, and S5‐S8. In five others one component was identified unambiguously. No additional complexes were seen when the longer homologous butyro and capro reagents were used