The Use of a Cleavable Crosslinking Reagent to Identify Neighboring Proteins in the 30‐S Ribosomal Subunit of Escherichia coli
- 1 March 1976
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 63 (1), 83-92
- https://doi.org/10.1111/j.1432-1033.1976.tb10210.x
Abstract
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 usedKeywords
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