Cargando…

Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity

[Image: see text] Bacterial glycomes are rich in prokaryote-specific or “rare” sugars that are absent in mammals. Like common sugars found across organisms, rare sugars are typically activated as nucleoside diphosphate sugars (NDP-sugars) by nucleotidyltransferases. In bacteria, the nucleotidyltrans...

Descripción completa

Detalles Bibliográficos
Autores principales: Zheng, Meng, Zheng, Maggie C., Kim, Hanee, Lupoli, Tania J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2023
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10375476/
https://www.ncbi.nlm.nih.gov/pubmed/37283497
http://dx.doi.org/10.1021/jacs.3c02319
_version_ 1785079044940759040
author Zheng, Meng
Zheng, Maggie C.
Kim, Hanee
Lupoli, Tania J.
author_facet Zheng, Meng
Zheng, Maggie C.
Kim, Hanee
Lupoli, Tania J.
author_sort Zheng, Meng
collection PubMed
description [Image: see text] Bacterial glycomes are rich in prokaryote-specific or “rare” sugars that are absent in mammals. Like common sugars found across organisms, rare sugars are typically activated as nucleoside diphosphate sugars (NDP-sugars) by nucleotidyltransferases. In bacteria, the nucleotidyltransferase RmlA initiates the production of several rare NDP-sugars, which in turn regulate downstream glycan assembly through feedback inhibition of RmlA via binding to an allosteric site. In vitro, RmlA activates a range of common sugar-1-phosphates to produce NDP-sugars for biochemical and synthetic applications. However, our ability to probe bacterial glycan biosynthesis is hindered by limited chemoenzymatic access to rare NDP-sugars. We postulate that natural feedback mechanisms impact nucleotidyltransferase utility. Here, we use synthetic rare NDP-sugars to identify structural features required for regulation of RmlA from diverse bacterial species. We find that mutation of RmlA to eliminate allosteric binding of an abundant rare NDP-sugar facilitates the activation of noncanonical rare sugar-1-phosphate substrates, as products no longer affect turnover. In addition to promoting an understanding of nucleotidyltransferase regulation by metabolites, this work provides new routes to access rare sugar substrates for the study of important bacteria-specific glycan pathways.
format Online
Article
Text
id pubmed-10375476
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-103754762023-07-29 Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity Zheng, Meng Zheng, Maggie C. Kim, Hanee Lupoli, Tania J. J Am Chem Soc [Image: see text] Bacterial glycomes are rich in prokaryote-specific or “rare” sugars that are absent in mammals. Like common sugars found across organisms, rare sugars are typically activated as nucleoside diphosphate sugars (NDP-sugars) by nucleotidyltransferases. In bacteria, the nucleotidyltransferase RmlA initiates the production of several rare NDP-sugars, which in turn regulate downstream glycan assembly through feedback inhibition of RmlA via binding to an allosteric site. In vitro, RmlA activates a range of common sugar-1-phosphates to produce NDP-sugars for biochemical and synthetic applications. However, our ability to probe bacterial glycan biosynthesis is hindered by limited chemoenzymatic access to rare NDP-sugars. We postulate that natural feedback mechanisms impact nucleotidyltransferase utility. Here, we use synthetic rare NDP-sugars to identify structural features required for regulation of RmlA from diverse bacterial species. We find that mutation of RmlA to eliminate allosteric binding of an abundant rare NDP-sugar facilitates the activation of noncanonical rare sugar-1-phosphate substrates, as products no longer affect turnover. In addition to promoting an understanding of nucleotidyltransferase regulation by metabolites, this work provides new routes to access rare sugar substrates for the study of important bacteria-specific glycan pathways. American Chemical Society 2023-06-07 /pmc/articles/PMC10375476/ /pubmed/37283497 http://dx.doi.org/10.1021/jacs.3c02319 Text en © 2023 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Zheng, Meng
Zheng, Maggie C.
Kim, Hanee
Lupoli, Tania J.
Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title_full Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title_fullStr Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title_full_unstemmed Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title_short Feedback Inhibition of Bacterial Nucleotidyltransferases by Rare Nucleotide l-Sugars Restricts Substrate Promiscuity
title_sort feedback inhibition of bacterial nucleotidyltransferases by rare nucleotide l-sugars restricts substrate promiscuity
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10375476/
https://www.ncbi.nlm.nih.gov/pubmed/37283497
http://dx.doi.org/10.1021/jacs.3c02319
work_keys_str_mv AT zhengmeng feedbackinhibitionofbacterialnucleotidyltransferasesbyrarenucleotidelsugarsrestrictssubstratepromiscuity
AT zhengmaggiec feedbackinhibitionofbacterialnucleotidyltransferasesbyrarenucleotidelsugarsrestrictssubstratepromiscuity
AT kimhanee feedbackinhibitionofbacterialnucleotidyltransferasesbyrarenucleotidelsugarsrestrictssubstratepromiscuity
AT lupolitaniaj feedbackinhibitionofbacterialnucleotidyltransferasesbyrarenucleotidelsugarsrestrictssubstratepromiscuity