Hybrid Mass Spectrometry Approaches to Determine How L-Histidine Feedback Regulates the Enzyzme MtATP-Phosphoribosyltransferase

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Title: Hybrid Mass Spectrometry Approaches to Determine How L-Histidine Feedback Regulates the Enzyzme MtATP-Phosphoribosyltransferase
Author(s): Pacholarz, KJ
Burnley, RJ
Jowitt, TA
Ordsmith, V
Pisco, JP
Porrini, M
Larrouy-Maumus, G
Garlish, RA
Taylor, RJ
De Carvalho, LPS
Barran, PE
Item Type: Journal Article
Abstract: MtATP-phosphoribosyltransferase (MtATP-PRT) is an enzyme catalyzing the first step of the biosynthesis of L-histidine in Mycobacterium tuberculosis, and proposed to be regulated via an allosteric mechanism. Native mass spectrometry (MS) reveals MtATP-PRT to exist as a hexamer. Conformational changes induced by L-histidine binding and the influence of buffer pH are determined with ion mobility MS, hydrogen deuterium exchange (HDX) MS, and analytical ultracentrifugation. The experimental collision cross-section (DTCCSHe) decreases from 76.6 to 73.5 nm2 upon ligand binding at pH 6.8, which correlates to the decrease in CCS calculated from crystal structures. No such changes in conformation were found at pH 9.0. Further detail on the regions that exhibit conformational change on L-histidine binding is obtained with HDX-MS experiments. On incubation with L-histidine, rapid changes are observed within domain III, and around the active site at longer times, indicating an allosteric effect.
Publication Date: 6-Apr-2017
Date of Acceptance: 9-Mar-2017
URI: http://hdl.handle.net/10044/1/48874
DOI: https://dx.doi.org/10.1016/j.str.2017.03.005
ISSN: 1878-4186
Publisher: Elsevier (Cell Press)
Start Page: 730
End Page: 738.e4
Journal / Book Title: Structure
Volume: 25
Issue: 5
Copyright Statement: © 2017 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
Keywords: Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Biophysics
Cell Biology
INDUCED CONFORMATIONAL-CHANGES
PROTEIN-LIGAND INTERACTIONS
STRUCTURAL BIOLOGY
HYDROGEN-EXCHANGE
ATP PHOSPHORIBOSYLTRANSFERASE
DRUG DISCOVERY
DYNAMICS
EQUATION
BINDING
ULTRACENTRIFUGATION
ATP-phosphoribosyltransferase
allostery
hydrogen deuterium exchange
ion mobility
protein conformation
structural mass spectrometry
tuberculosis
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Biophysics
Cell Biology
INDUCED CONFORMATIONAL-CHANGES
PROTEIN-LIGAND INTERACTIONS
STRUCTURAL BIOLOGY
HYDROGEN-EXCHANGE
ATP PHOSPHORIBOSYLTRANSFERASE
DRUG DISCOVERY
DYNAMICS
EQUATION
BINDING
ULTRACENTRIFUGATION
Biophysics
06 Biological Sciences
08 Information And Computing Sciences
03 Chemical Sciences
Publication Status: Published
Appears in Collections:Faculty of Natural Sciences



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