Cargando…

Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451

The main enzymatic reaction of the large ribosomal subunit is peptide bond formation. Ribosome crystallography showed that A2451 of 23S rRNA makes the closest approach to the attacking amino group of aminoacyl-tRNA. Mutations of A2451 had relatively small effects on transpeptidation and failed to un...

Descripción completa

Detalles Bibliográficos
Autores principales: Erlacher, Matthias D., Lang, Kathrin, Shankaran, Nisha, Wotzel, Brigitte, Hüttenhofer, Alexander, Micura, Ronald, Mankin, Alexander S., Polacek, Norbert
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2005
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1065261/
https://www.ncbi.nlm.nih.gov/pubmed/15767286
http://dx.doi.org/10.1093/nar/gki308
_version_ 1782123358258724864
author Erlacher, Matthias D.
Lang, Kathrin
Shankaran, Nisha
Wotzel, Brigitte
Hüttenhofer, Alexander
Micura, Ronald
Mankin, Alexander S.
Polacek, Norbert
author_facet Erlacher, Matthias D.
Lang, Kathrin
Shankaran, Nisha
Wotzel, Brigitte
Hüttenhofer, Alexander
Micura, Ronald
Mankin, Alexander S.
Polacek, Norbert
author_sort Erlacher, Matthias D.
collection PubMed
description The main enzymatic reaction of the large ribosomal subunit is peptide bond formation. Ribosome crystallography showed that A2451 of 23S rRNA makes the closest approach to the attacking amino group of aminoacyl-tRNA. Mutations of A2451 had relatively small effects on transpeptidation and failed to unequivocally identify the crucial functional group(s). Here, we employed an in vitro reconstitution system for chemical engineering the peptidyl transferase center by introducing non-natural nucleosides at position A2451. This allowed us to investigate the peptidyl transfer reaction performed by a ribosome that contained a modified nucleoside at the active site. The main finding is that ribosomes carrying a 2′-deoxyribose at A2451 showed a compromised peptidyl transferase activity. In variance, adenine base modifications and even the removal of the entire nucleobase at A2451 had only little impact on peptide bond formation, as long as the 2′-hydroxyl was present. This implicates a functional or structural role of the 2′-hydroxyl group at A2451 for transpeptidation.
format Text
id pubmed-1065261
institution National Center for Biotechnology Information
language English
publishDate 2005
publisher Oxford University Press
record_format MEDLINE/PubMed
spelling pubmed-10652612005-03-15 Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451 Erlacher, Matthias D. Lang, Kathrin Shankaran, Nisha Wotzel, Brigitte Hüttenhofer, Alexander Micura, Ronald Mankin, Alexander S. Polacek, Norbert Nucleic Acids Res Article The main enzymatic reaction of the large ribosomal subunit is peptide bond formation. Ribosome crystallography showed that A2451 of 23S rRNA makes the closest approach to the attacking amino group of aminoacyl-tRNA. Mutations of A2451 had relatively small effects on transpeptidation and failed to unequivocally identify the crucial functional group(s). Here, we employed an in vitro reconstitution system for chemical engineering the peptidyl transferase center by introducing non-natural nucleosides at position A2451. This allowed us to investigate the peptidyl transfer reaction performed by a ribosome that contained a modified nucleoside at the active site. The main finding is that ribosomes carrying a 2′-deoxyribose at A2451 showed a compromised peptidyl transferase activity. In variance, adenine base modifications and even the removal of the entire nucleobase at A2451 had only little impact on peptide bond formation, as long as the 2′-hydroxyl was present. This implicates a functional or structural role of the 2′-hydroxyl group at A2451 for transpeptidation. Oxford University Press 2005 2005-03-14 /pmc/articles/PMC1065261/ /pubmed/15767286 http://dx.doi.org/10.1093/nar/gki308 Text en © The Author 2005. Published by Oxford University Press. All rights reserved
spellingShingle Article
Erlacher, Matthias D.
Lang, Kathrin
Shankaran, Nisha
Wotzel, Brigitte
Hüttenhofer, Alexander
Micura, Ronald
Mankin, Alexander S.
Polacek, Norbert
Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title_full Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title_fullStr Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title_full_unstemmed Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title_short Chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of A2451
title_sort chemical engineering of the peptidyl transferase center reveals an important role of the 2′-hydroxyl group of a2451
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC1065261/
https://www.ncbi.nlm.nih.gov/pubmed/15767286
http://dx.doi.org/10.1093/nar/gki308
work_keys_str_mv AT erlachermatthiasd chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT langkathrin chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT shankarannisha chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT wotzelbrigitte chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT huttenhoferalexander chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT micuraronald chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT mankinalexanders chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451
AT polaceknorbert chemicalengineeringofthepeptidyltransferasecenterrevealsanimportantroleofthe2hydroxylgroupofa2451