Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli

Abstract Metabolite binding to proteins regulates nearly all cellular processes, but our knowledge of these interactions originates primarily from empirical in vitro studies. Here, we report the first systematic study of interactions between water‐soluble proteins and polar metabolites in an entire...

Full description

Saved in:
Bibliographic Details
Main Authors: Maren Diether, Yaroslav Nikolaev, Frédéric HT Allain, Uwe Sauer
Format: Article
Language:English
Published: Springer Nature 2019-08-01
Series:Molecular Systems Biology
Subjects:
Online Access:https://doi.org/10.15252/msb.20199008
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1849235289031049216
author Maren Diether
Yaroslav Nikolaev
Frédéric HT Allain
Uwe Sauer
author_facet Maren Diether
Yaroslav Nikolaev
Frédéric HT Allain
Uwe Sauer
author_sort Maren Diether
collection DOAJ
description Abstract Metabolite binding to proteins regulates nearly all cellular processes, but our knowledge of these interactions originates primarily from empirical in vitro studies. Here, we report the first systematic study of interactions between water‐soluble proteins and polar metabolites in an entire biological subnetwork. To test the depth of our current knowledge, we chose to investigate the well‐characterized Escherichia coli central metabolism. Using ligand‐detected NMR, we assayed 29 enzymes towards binding events with 55 intracellular metabolites. Focusing on high‐confidence interactions at a false‐positive rate of 5%, we detected 98 interactions, among which purine nucleotides accounted for one‐third, while 50% of all metabolites did not interact with any enzyme. In contrast, only five enzymes did not exhibit any metabolite binding and some interacted with up to 11 metabolites. About 40% of the interacting metabolites were predicted to be allosteric effectors based on low chemical similarity to their target's reactants. For five of the eight tested interactions, in vitro assays confirmed novel regulatory functions, including ATP and GTP inhibition of the first pentose phosphate pathway enzyme. With 76 new candidate regulatory interactions that have not been reported previously, we essentially doubled the number of known interactions, indicating that the presently available information about protein–metabolite interactions may only be the tip of the iceberg.
format Article
id doaj-art-4586648bbd6e4aae9d16bfd20a2de217
institution Kabale University
issn 1744-4292
language English
publishDate 2019-08-01
publisher Springer Nature
record_format Article
series Molecular Systems Biology
spelling doaj-art-4586648bbd6e4aae9d16bfd20a2de2172025-08-20T04:02:50ZengSpringer NatureMolecular Systems Biology1744-42922019-08-0115811610.15252/msb.20199008Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coliMaren Diether0Yaroslav Nikolaev1Frédéric HT Allain2Uwe Sauer3Institute of Molecular Systems Biology, ETH ZurichInstitute of Molecular Biology and Biophysics, ETH ZurichInstitute of Molecular Biology and Biophysics, ETH ZurichInstitute of Molecular Systems Biology, ETH ZurichAbstract Metabolite binding to proteins regulates nearly all cellular processes, but our knowledge of these interactions originates primarily from empirical in vitro studies. Here, we report the first systematic study of interactions between water‐soluble proteins and polar metabolites in an entire biological subnetwork. To test the depth of our current knowledge, we chose to investigate the well‐characterized Escherichia coli central metabolism. Using ligand‐detected NMR, we assayed 29 enzymes towards binding events with 55 intracellular metabolites. Focusing on high‐confidence interactions at a false‐positive rate of 5%, we detected 98 interactions, among which purine nucleotides accounted for one‐third, while 50% of all metabolites did not interact with any enzyme. In contrast, only five enzymes did not exhibit any metabolite binding and some interacted with up to 11 metabolites. About 40% of the interacting metabolites were predicted to be allosteric effectors based on low chemical similarity to their target's reactants. For five of the eight tested interactions, in vitro assays confirmed novel regulatory functions, including ATP and GTP inhibition of the first pentose phosphate pathway enzyme. With 76 new candidate regulatory interactions that have not been reported previously, we essentially doubled the number of known interactions, indicating that the presently available information about protein–metabolite interactions may only be the tip of the iceberg.https://doi.org/10.15252/msb.20199008allosterycentral metabolismnuclear magnetic resonanceprotein–metabolite interactionsregulation
spellingShingle Maren Diether
Yaroslav Nikolaev
Frédéric HT Allain
Uwe Sauer
Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
Molecular Systems Biology
allostery
central metabolism
nuclear magnetic resonance
protein–metabolite interactions
regulation
title Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
title_full Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
title_fullStr Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
title_full_unstemmed Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
title_short Systematic mapping of protein‐metabolite interactions in central metabolism of Escherichia coli
title_sort systematic mapping of protein metabolite interactions in central metabolism of escherichia coli
topic allostery
central metabolism
nuclear magnetic resonance
protein–metabolite interactions
regulation
url https://doi.org/10.15252/msb.20199008
work_keys_str_mv AT marendiether systematicmappingofproteinmetaboliteinteractionsincentralmetabolismofescherichiacoli
AT yaroslavnikolaev systematicmappingofproteinmetaboliteinteractionsincentralmetabolismofescherichiacoli
AT frederichtallain systematicmappingofproteinmetaboliteinteractionsincentralmetabolismofescherichiacoli
AT uwesauer systematicmappingofproteinmetaboliteinteractionsincentralmetabolismofescherichiacoli