Cross-linking of selected residues in the N- and C-terminal domains of Escherichia coli protein L7/L12 to other ribosomal proteins and the effect of elongation factor Tu.

The Journal of Biological Chemistry
D DeyR R Traut

Abstract

Five different variants of protein L7/L12, each with a single cysteine substitution at a selected site, were produced, modified with 125I-N-[4-(p-azidosalicylamido)-butyl]-3-(2'-pyridyldithio)propion amide, a radiolabeled, sulfhydryl-specific, heterobifunctional, cleavable photocross-linking reagent that transfers radiolabel to the target molecule upon reduction of the disulfide bond. The proteins were reconstituted with core particles depleted of wild type L7/L12 to yield 70 S ribosomes. Cross-linked molecules were identified and quantified by the radiolabel. No cross-linking of RNA was detected. Two sites in the dimeric N-terminal domain, Cys-12 and Cys-33, cross-linked strongly to L10 and in lower yield to L11 but to no other proteins. The three sites in the globular C-terminal domain all cross-linked strongly to L11 and, in lower yield, to L10. Weaker cross-linking to 50 S proteins L2 and L5 occurred from all three C-terminal domain locations. The 30 S ribosomal proteins S2, S3, S7, S14, S18 were also cross-linked from all three of these sites. Binding of the ternary complex [14C]Phe-tRNA-elongation factor Tu.guanyl-5'-yl imidodiphosphate) but not [14C]Phe-tRNA.elongation factor Tu.GDP.kirromycin increased labeling of L2, L...Continue Reading

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Citations

Dec 14, 2002·Biochemical and Biophysical Research Communications·Dmitri A KazminJean R Starkey
Dec 15, 2000·Current Opinion in Structural Biology·S Chandra Sanyal, A Liljas
May 31, 2011·Journal of Proteome Research·Matthew A Lauber, James P Reilly
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Nov 6, 2007·Journal of Bacteriology·Catherine Wicker-PlanquartJean-Michel Jault
Jul 21, 2004·Proceedings of the National Academy of Sciences of the United States of America·John ChristodoulouChristopher M Dobson
Jun 3, 2008·Journal of Molecular Biology·Yuliya GordiyenkoCarol V Robinson
Oct 1, 2010·Advanced Materials·Sonja BauhuberAchim Göpferich
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Feb 13, 2004·The Journal of Biological Chemistry·Eduard V BocharovAlexander S Arseniev
Jan 1, 2018·Quarterly Reviews of Biophysics·Anders Liljas, Suparna Sanyal

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