Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa

Cameron Baines; Jacob Sargeant; Christopher D. Fage; Hannah Pugh; Lona M. Alkhalaf; Gregory L. Challis; Neil J. Oldham

doi:10.1021/jasms.4c00087

Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa

Cameron Baines
, Jacob Sargeant
, Christopher D. Fage
, Hannah Pugh
, Lona M. Alkhalaf
, Gregory L. Challis
, Neil J. Oldham

Research output: Contribution to journal › Article › Research › peer-review

1 Citation (Scopus)

Abstract

Collision-induced unfolding (CIU) of protein ions, monitored by ion mobility-mass spectrometry, can be used to assess the stability of their compact gas-phase fold and hence provide structural information. The bacterial elongation factor EF-Tu, a key protein for mRNA translation in prokaryotes and hence a promising antibiotic target, has been studied by CIU. The major [M + 12H]¹²⁺ ion of EF-Tu unfolded in collision with Ar atoms between 40 and 50 V, corresponding to an E_lab energy of 480-500 eV. Binding of the cofactor analogue GDPNP and the antibiotic enacyloxin IIa stabilized the compact fold of EF-Tu, although dissociation of the latter from the complex diminished its stabilizing effect at higher collision energies. Molecular dynamics simulations of the [M + 12H]¹²⁺ EF-Tu ion showed similar qualitative behavior to the experimental results.

Original language	English
Pages (from-to)	1490-1496
Number of pages	7
Journal	Journal of the American Society for Mass Spectrometry
Volume	35
Issue number	7
DOIs	https://doi.org/10.1021/jasms.4c00087
Publication status	Published - 3 Jul 2024

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10.1021/jasms.4c00087Licence: CC BY

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@article{d559b6a5544242bcb72a1c569c15a776,

title = "Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa",

abstract = "Collision-induced unfolding (CIU) of protein ions, monitored by ion mobility-mass spectrometry, can be used to assess the stability of their compact gas-phase fold and hence provide structural information. The bacterial elongation factor EF-Tu, a key protein for mRNA translation in prokaryotes and hence a promising antibiotic target, has been studied by CIU. The major [M + 12H]12+ ion of EF-Tu unfolded in collision with Ar atoms between 40 and 50 V, corresponding to an Elab energy of 480-500 eV. Binding of the cofactor analogue GDPNP and the antibiotic enacyloxin IIa stabilized the compact fold of EF-Tu, although dissociation of the latter from the complex diminished its stabilizing effect at higher collision energies. Molecular dynamics simulations of the [M + 12H]12+ EF-Tu ion showed similar qualitative behavior to the experimental results.",

author = "Cameron Baines and Jacob Sargeant and Fage, \{Christopher D.\} and Hannah Pugh and Alkhalaf, \{Lona M.\} and Challis, \{Gregory L.\} and Oldham, \{Neil J.\}",

note = "Funding Information: ChargePlacer alongside other Python scripts used during this work were developed by Jedd Bellamy-Carter (Loughborough University). Protein structures were visualized in UCSF ChimeraX (Resource for Biocomputing, Visualization and Informatics at the University of California, San Francisco). Data analyses were performed in GraphPad Prism 10.0.2, SideFX Houdini 19.5, and VMD (Theoretical and Computational Biophysics Group, University of Illinois). C.B. was supported in full by the UKRI Biotechnology and Biological Sciences Research Council (grant BB/T008369/1). J.S. was supported by a National Productivity Investment Fund grant from the BBSRC (BB/R505845/1 to G.L.C.). C.D.F. was supported by a research grant from the BBSRC (BB/R010218/1 to G.L.C.). H.P. was supported by the BBSRC-funded Midlands Integrative Bioscience Doctoral Training Partnership (BB/J014532/1). Publisher Copyright: {\textcopyright} 2024 The Authors. Published by American Chemical Society.",

year = "2024",

month = jul,

day = "3",

doi = "10.1021/jasms.4c00087",

language = "English",

volume = "35",

pages = "1490--1496",

journal = "Journal of the American Society for Mass Spectrometry",

issn = "1044-0305",

publisher = "Springer-Verlag London Ltd.",

number = "7",

}

Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa. / Baines, Cameron; Sargeant, Jacob; Fage, Christopher D. et al.
In: Journal of the American Society for Mass Spectrometry, Vol. 35, No. 7, 03.07.2024, p. 1490-1496.

Research output: Contribution to journal › Article › Research › peer-review

TY - JOUR

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AU - Pugh, Hannah

AU - Alkhalaf, Lona M.

AU - Challis, Gregory L.

AU - Oldham, Neil J.

N1 - Funding Information: ChargePlacer alongside other Python scripts used during this work were developed by Jedd Bellamy-Carter (Loughborough University). Protein structures were visualized in UCSF ChimeraX (Resource for Biocomputing, Visualization and Informatics at the University of California, San Francisco). Data analyses were performed in GraphPad Prism 10.0.2, SideFX Houdini 19.5, and VMD (Theoretical and Computational Biophysics Group, University of Illinois). C.B. was supported in full by the UKRI Biotechnology and Biological Sciences Research Council (grant BB/T008369/1). J.S. was supported by a National Productivity Investment Fund grant from the BBSRC (BB/R505845/1 to G.L.C.). C.D.F. was supported by a research grant from the BBSRC (BB/R010218/1 to G.L.C.). H.P. was supported by the BBSRC-funded Midlands Integrative Bioscience Doctoral Training Partnership (BB/J014532/1). Publisher Copyright: © 2024 The Authors. Published by American Chemical Society.

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N2 - Collision-induced unfolding (CIU) of protein ions, monitored by ion mobility-mass spectrometry, can be used to assess the stability of their compact gas-phase fold and hence provide structural information. The bacterial elongation factor EF-Tu, a key protein for mRNA translation in prokaryotes and hence a promising antibiotic target, has been studied by CIU. The major [M + 12H]12+ ion of EF-Tu unfolded in collision with Ar atoms between 40 and 50 V, corresponding to an Elab energy of 480-500 eV. Binding of the cofactor analogue GDPNP and the antibiotic enacyloxin IIa stabilized the compact fold of EF-Tu, although dissociation of the latter from the complex diminished its stabilizing effect at higher collision energies. Molecular dynamics simulations of the [M + 12H]12+ EF-Tu ion showed similar qualitative behavior to the experimental results.

AB - Collision-induced unfolding (CIU) of protein ions, monitored by ion mobility-mass spectrometry, can be used to assess the stability of their compact gas-phase fold and hence provide structural information. The bacterial elongation factor EF-Tu, a key protein for mRNA translation in prokaryotes and hence a promising antibiotic target, has been studied by CIU. The major [M + 12H]12+ ion of EF-Tu unfolded in collision with Ar atoms between 40 and 50 V, corresponding to an Elab energy of 480-500 eV. Binding of the cofactor analogue GDPNP and the antibiotic enacyloxin IIa stabilized the compact fold of EF-Tu, although dissociation of the latter from the complex diminished its stabilizing effect at higher collision energies. Molecular dynamics simulations of the [M + 12H]12+ EF-Tu ion showed similar qualitative behavior to the experimental results.

UR - https://www.scopus.com/pages/publications/85195263503

U2 - 10.1021/jasms.4c00087

DO - 10.1021/jasms.4c00087

M3 - Article

C2 - 38830009

AN - SCOPUS:85195263503

SN - 1044-0305

VL - 35

SP - 1490

EP - 1496

JO - Journal of the American Society for Mass Spectrometry

JF - Journal of the American Society for Mass Spectrometry

IS - 7

ER -

Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa

Abstract

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ARC Centre of Excellence for Innovations in Peptide and Protein Science

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Native ESI-MS and Collision-Induced Unfolding (CIU) of the Complex between Bacterial Elongation Factor-Tu and the Antibiotic Enacyloxin IIa

Abstract

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ARC Centre of Excellence for Innovations in Peptide and Protein Science

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