Native Ambient Mass Spectrometry Enables Analysis of Intact Endogenous Protein Assemblies up to 145 kDa Directly from Tissue

Oliver J Hale; James W Hughes; Emma K Sisley; Helen J Cooper

doi:10.1021/acs.analchem.1c05353

Native Ambient Mass Spectrometry Enables Analysis of Intact Endogenous Protein Assemblies up to 145 kDa Directly from Tissue

Oliver J Hale, James W Hughes, Emma K Sisley, Helen J Cooper

Biosciences

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Abstract

Untargeted label-free interrogation of proteins in their functional form directly from their physiological environment promises to transform life sciences research by providing unprecedented insight into their transient interactions with other biomolecules and xenobiotics. Native ambient mass spectrometry (NAMS) shows great potential for the structural analysis of endogenous protein assemblies directly from tissues; however, to date, this has been limited to assemblies of low molecular weight (<20 kDa) or very high abundance (hemoglobin tetramer in blood vessels, RidA homotrimer in kidney cortex tissues). The present work constitutes a step change for NAMS of protein assemblies: we demonstrate the detection and identification of a range of intact endogenous protein assemblies with various stoichiometries (dimer, trimer, and tetramer) from a range of tissue types (brain, kidney, liver) by the use of multiple NAMS techniques. Crucially, we demonstrate a greater than twofold increase in accessible molecular weight (up to 145 kDa). In addition, spatial distributions of protein assemblies up to 94 kDa were mapped in brain and kidney by nanospray desorption electrospray ionization (nano-DESI) mass spectrometry imaging.

Original language	English
Pages (from-to)	5608-5614
Number of pages	7
Journal	Analytical Chemistry
Volume	94
Issue number	14
Early online date	31 Mar 2022
DOIs	https://doi.org/10.1021/acs.analchem.1c05353
Publication status	Published - 12 Apr 2022

Bibliographical note

Funding Information:
H.J.C. is an EPSRC Established Career Fellow (EP/S002979/1). O.J.H. and J.W.H. were funded by EPSRC (EP/S002979/1). E.K.S. was funded by the University of Birmingham. The Orbitrap Eclipse mass spectrometer used in this work was funded by BBSRC (BB/S019456/1). The Q-Exactive HF mass spectrometer was funded by the University of Birmingham. The authors thank Thermo Fisher Scientific for access to modified software that enabled native MS analysis on the Q-Exactive HF mass spectrometer. Funding for the equipment used to make the nanoelectrospray emitters was provided by The Royal Society (RGS\R1\201411). Supplementary data supporting this research is openly available from https://doi.org/10.25500/edata.bham.00000797 .

Publisher Copyright:
© 2022 American Chemical Society. All rights reserved.

Keywords

Animals
Brain/metabolism
Kidney/metabolism
Proteins/metabolism
Scrapie
Sheep
Spectrometry, Mass, Electrospray Ionization/methods

ASJC Scopus subject areas

Analytical Chemistry

Access to Document

10.1021/acs.analchem.1c05353Licence: Creative Commons: Attribution (CC BY)

acs.analchem.1c05353Final published version, 3.72 MBLicence: Creative Commons: Attribution (CC BY)

A new mass spectrometer for structural proteomics and protein imaging
Thomas, C., Grant, M., Lovering, A., Alderwick, L., Tomlinson, M., Winn, P., Knowles, T., Cooper, H., Styles, I., Gibbs, D., Huber, D., Bhatt, A. & Leney, A.
Biotechnology & Biological Sciences Research Council
1/07/19 → 30/06/20
Project: Research
NAMS: Native ambient mass spectrometry
Cooper, H.
Engineering & Physical Science Research Council
1/06/19 → 30/11/23
Project: Research

Cite this

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title = "Native Ambient Mass Spectrometry Enables Analysis of Intact Endogenous Protein Assemblies up to 145 kDa Directly from Tissue",

abstract = "Untargeted label-free interrogation of proteins in their functional form directly from their physiological environment promises to transform life sciences research by providing unprecedented insight into their transient interactions with other biomolecules and xenobiotics. Native ambient mass spectrometry (NAMS) shows great potential for the structural analysis of endogenous protein assemblies directly from tissues; however, to date, this has been limited to assemblies of low molecular weight (<20 kDa) or very high abundance (hemoglobin tetramer in blood vessels, RidA homotrimer in kidney cortex tissues). The present work constitutes a step change for NAMS of protein assemblies: we demonstrate the detection and identification of a range of intact endogenous protein assemblies with various stoichiometries (dimer, trimer, and tetramer) from a range of tissue types (brain, kidney, liver) by the use of multiple NAMS techniques. Crucially, we demonstrate a greater than twofold increase in accessible molecular weight (up to 145 kDa). In addition, spatial distributions of protein assemblies up to 94 kDa were mapped in brain and kidney by nanospray desorption electrospray ionization (nano-DESI) mass spectrometry imaging.",

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note = "Funding Information: H.J.C. is an EPSRC Established Career Fellow (EP/S002979/1). O.J.H. and J.W.H. were funded by EPSRC (EP/S002979/1). E.K.S. was funded by the University of Birmingham. The Orbitrap Eclipse mass spectrometer used in this work was funded by BBSRC (BB/S019456/1). The Q-Exactive HF mass spectrometer was funded by the University of Birmingham. The authors thank Thermo Fisher Scientific for access to modified software that enabled native MS analysis on the Q-Exactive HF mass spectrometer. Funding for the equipment used to make the nanoelectrospray emitters was provided by The Royal Society (RGS\R1\201411). Supplementary data supporting this research is openly available from https://doi.org/10.25500/edata.bham.00000797 . Publisher Copyright: {\textcopyright} 2022 American Chemical Society. All rights reserved.",

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N2 - Untargeted label-free interrogation of proteins in their functional form directly from their physiological environment promises to transform life sciences research by providing unprecedented insight into their transient interactions with other biomolecules and xenobiotics. Native ambient mass spectrometry (NAMS) shows great potential for the structural analysis of endogenous protein assemblies directly from tissues; however, to date, this has been limited to assemblies of low molecular weight (<20 kDa) or very high abundance (hemoglobin tetramer in blood vessels, RidA homotrimer in kidney cortex tissues). The present work constitutes a step change for NAMS of protein assemblies: we demonstrate the detection and identification of a range of intact endogenous protein assemblies with various stoichiometries (dimer, trimer, and tetramer) from a range of tissue types (brain, kidney, liver) by the use of multiple NAMS techniques. Crucially, we demonstrate a greater than twofold increase in accessible molecular weight (up to 145 kDa). In addition, spatial distributions of protein assemblies up to 94 kDa were mapped in brain and kidney by nanospray desorption electrospray ionization (nano-DESI) mass spectrometry imaging.

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Native Ambient Mass Spectrometry Enables Analysis of Intact Endogenous Protein Assemblies up to 145 kDa Directly from Tissue

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

Access to Document

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Projects

A new mass spectrometer for structural proteomics and protein imaging

NAMS: Native ambient mass spectrometry

Cite this