article · 16/08/2017
Affinity labelling in situ of the bL12 protein on E. coli 70S ribosomes by means of a tRNA dialdehyde derivative
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Résumé
In this report, we have used periodate-oxidized tRNA (tRNAox) as an affinity laleling reagent to demonstrate that: (i) the bL12 protein contacts the CCA-arm of P- site bound tRNA on the Escherichia coli 70S ribo- somes; (ii) the stoichiometry of labelling is one molecule of tRNAox bound to one polypeptide chain of endogen- ous bL12; (iii) cross-linking in situ of bL12 with tRNAox on the ribosomes provokes the loss of activity; (iv) intact tRNA protects bL12 in the 70S ribosomes against cross-linking with tRNAox; (v) both tRNAox and pyridoxal 50-phosphate (PLP) compete for the same or for proximal cross-linking site(s) on bL12 inside the ribosome; (vi) the stoichiometry of cross-link- ing of PLP to the recombinant E. coli bL12 protein is one molecule of PLP covalently bound per polypeptide chain; (vii) the amino acid residue of recombinant bL12 cross-linked with PLP is Lys-65; (viii) Lys-65 of E. coli bL12 corresponds to Lys-53 of eL42 which was previ- ously shown to cross-link with P-site bound tRNAox on human 80S ribosomes in situ; (ix) finally, E. coli bL12 and human eL42 proteins display significant primary structure similarities, which argues for evolutionary conservation of these two proteins located at the tRNA-CCA binding site on eubacterial and eukaryal ribosomes.
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Hountondji, C., Crechet, J.-B., Le Caër, J.-P., Lancelot, V., Cognet, J.-A.-H., & Baouz, S. (2017). Affinity labelling in situ of the bL12 protein on E. coli 70S ribosomes by means of a tRNA dialdehyde derivative. J. Biochem., 162(6), 437-448. https://doi.org/doi:10.1093/jb/mvx055@article{Hountondji2017_163,
author = {Hountondji, Codjo and Crechet, Jean-Bernard and Le Caër, Jean-Pierre and Lancelot, Véronique and Cognet, Jean A. H. and Baouz, Soria},
year = {2017},
month = {8},
title = {Affinity labelling in situ of the bL12 protein on E. coli 70S ribosomes by means of a tRNA dialdehyde derivative},
journal = {J. Biochem.},
editor = {Oxford University Press},
volume = {162},
number = {6},
pages = {437-448},
abstract = {In this report, we have used periodate-oxidized tRNA (tRNAox) as an affinity laleling reagent to demonstrate that: (i) the bL12 protein contacts the CCA-arm of P- site bound tRNA on the Escherichia coli 70S ribo- somes; (ii) the stoichiometry of labelling is one molecule of tRNAox bound to one polypeptide chain of endogen- ous bL12; (iii) cross-linking in situ of bL12 with tRNAox on the ribosomes provokes the loss of activity; (iv) intact tRNA protects bL12 in the 70S ribosomes against cross-linking with tRNAox; (v) both tRNAox and pyridoxal 50-phosphate (PLP) compete for the same or for proximal cross-linking site(s) on bL12 inside the ribosome; (vi) the stoichiometry of cross-link- ing of PLP to the recombinant E. coli bL12 protein is one molecule of PLP covalently bound per polypeptide chain; (vii) the amino acid residue of recombinant bL12 cross-linked with PLP is Lys-65; (viii) Lys-65 of E. coli bL12 corresponds to Lys-53 of eL42 which was previ- ously shown to cross-link with P-site bound tRNAox on human 80S ribosomes in situ; (ix) finally, E. coli bL12 and human eL42 proteins display significant primary structure similarities, which argues for evolutionary conservation of these two proteins located at the tRNA-CCA binding site on eubacterial and eukaryal ribosomes.},
url = {https://academic.oup.com/jb/article-abstract/doi/10.1093/jb/mvx055/4082913?redirectedFrom=fulltext},
doi = {doi:10.1093/jb/mvx055},
}TY - JOUR
AU - Hountondji, Codjo
AU - Crechet, Jean-Bernard
AU - Le Caër, Jean-Pierre
AU - Lancelot, Véronique
AU - Cognet, Jean A. H.
AU - Baouz, Soria
PY - 2017
DA - 2017/08/16
TI - Affinity labelling in situ of the bL12 protein on E. coli 70S ribosomes by means of a tRNA dialdehyde derivative
JO - J. Biochem.
VL - 162
IS - 6
AB - In this report, we have used periodate-oxidized tRNA (tRNAox) as an affinity laleling reagent to demonstrate that: (i) the bL12 protein contacts the CCA-arm of P- site bound tRNA on the Escherichia coli 70S ribo- somes; (ii) the stoichiometry of labelling is one molecule of tRNAox bound to one polypeptide chain of endogen- ous bL12; (iii) cross-linking in situ of bL12 with tRNAox on the ribosomes provokes the loss of activity; (iv) intact tRNA protects bL12 in the 70S ribosomes against cross-linking with tRNAox; (v) both tRNAox and pyridoxal 50-phosphate (PLP) compete for the same or for proximal cross-linking site(s) on bL12 inside the ribosome; (vi) the stoichiometry of cross-link- ing of PLP to the recombinant E. coli bL12 protein is one molecule of PLP covalently bound per polypeptide chain; (vii) the amino acid residue of recombinant bL12 cross-linked with PLP is Lys-65; (viii) Lys-65 of E. coli bL12 corresponds to Lys-53 of eL42 which was previ- ously shown to cross-link with P-site bound tRNAox on human 80S ribosomes in situ; (ix) finally, E. coli bL12 and human eL42 proteins display significant primary structure similarities, which argues for evolutionary conservation of these two proteins located at the tRNA-CCA binding site on eubacterial and eukaryal ribosomes.
SP - 437
EP - 448
DO - doi:10.1093/jb/mvx055
UR - https://academic.oup.com/jb/article-abstract/doi/10.1093/jb/mvx055/4082913?redirectedFrom=fulltext
ER -