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Peer-reviewedCC BYAnalytical Chemistry

Nanopipettes Enable Native Mass Spectrometry Studies of the Intrinsically Disordered Protein α-Synuclein in Biochemical Buffers

  • Emily J. Byrd1,2
  • Emma L. Norgate1,2
  • Joel A. Crossley1,2
  • Chalmers Chi Cheng Chau3,4,2
  • Bob Schiffrin1,2
  • Alexander N. Kulak5,2
  • Sheena E. Radford1,2
  • Paolo Actis3,4,2
  • Antonio N. Calabrese1,2
  • Frank Sobott1,2
5 affiliations
  1. 1Astbury Centre for Structural Molecular Biology, School of Molecular and Cellular Biology, Faculty of Biological Sciences
  2. 2University of Leeds
  3. 3Bragg Centre for Materials Research
  4. 4School of Electronic and Electrical Engineering
  5. 5School of Chemistry
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Abstract

Electrolyte conditions in vivo and in vitro are known to influence protein structure and function. Intrinsically disordered proteins (IDPs) are particularly sensitive to their solution conditions such as ionic strength and molecular crowding, and their dynamic structural ensembles rapidly respond to the cellular environment. While structural mass spectrometry (MS) techniques are uniquely able to capture aspects of this structural diversity, technical limitations have largely precluded the use of native MS approaches to interrogate the conformational rearrangements of IDPs in response to high concentrations of non-volatile salts. Here, we overcome this challenge by employing sub 100-nm nanopipette electrospray emitters for more gentle and salt-tolerant analysis to study the conformational distribution of α-Synuclein (αS) using native MS and ion mobility-MS in varied solution conditions, including in phosphate buffered saline. We show using native MS that it is possible to capture salt and buffer induced changes in the αS conformational ensemble when using traditional biochemical buffers, which reflect structural changes from in silico predictions and in-solution measurements. This work demonstrates the power of nanopipette emitters for the study of IDPs, and establishes native MS as a method that can be routinely used to determine how solution conditions tune the conformational landscape of IDPs.

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Cite

BibTeXRIS
BibTeX
@article{byrd2026nanopipettes,
  title = {{Nanopipettes Enable Native Mass Spectrometry Studies of the Intrinsically Disordered Protein α-Synuclein in Biochemical Buffers}},
  author = {Emily J. Byrd and Emma L. Norgate and Joel A. Crossley and Chalmers Chi Cheng Chau and Bob Schiffrin and Alexander N. Kulak and Sheena E. Radford and Paolo Actis and Antonio N. Calabrese and Frank Sobott},
  journal = {Analytical Chemistry},
  year = {2026},
  month = may,
  doi = {10.1021/acs.analchem.6c00544},
  url = {https://doi.org/10.1021/acs.analchem.6c00544},
  note = {CC BY},
}
RIS
TY  - JOUR
TI  - Nanopipettes Enable Native Mass Spectrometry Studies of the Intrinsically Disordered Protein α-Synuclein in Biochemical Buffers
AU  - Emily J. Byrd
AU  - Emma L. Norgate
AU  - Joel A. Crossley
AU  - Chalmers Chi Cheng Chau
AU  - Bob Schiffrin
AU  - Alexander N. Kulak
AU  - Sheena E. Radford
AU  - Paolo Actis
AU  - Antonio N. Calabrese
AU  - Frank Sobott
PY  - 2026
DA  - 2026/05/25
JO  - Analytical Chemistry
DO  - 10.1021/acs.analchem.6c00544
UR  - https://doi.org/10.1021/acs.analchem.6c00544
AB  - Electrolyte conditions in vivo and in vitro are known to influence protein structure and function. Intrinsically disordered proteins (IDPs) are particularly sensitive to their solution conditions such as ionic strength and molecular crowding, and their dynamic structural ensembles rapidly respond to the cellular environment. While structural mass spectrometry (MS) techniques are uniquely able to capture aspects of this structural diversity, technical limitations have largely precluded the use of native MS approaches to interrogate the conformational rearrangements of IDPs in response to high concentrations of non-volatile salts. Here, we overcome this challenge by employing sub 100-nm nanopipette electrospray emitters for more gentle and salt-tolerant analysis to study the conformational distribution of α-Synuclein (αS) using native MS and ion mobility-MS in varied solution conditions, including in phosphate buffered saline. We show using native MS that it is possible to capture salt and buffer induced changes in the αS conformational ensemble when using traditional biochemical buffers, which reflect structural changes from in silico predictions and in-solution measurements. This work demonstrates the power of nanopipette emitters for the study of IDPs, and establishes native MS as a method that can be routinely used to determine how solution conditions tune the conformational landscape of IDPs.
SN  - 0003-2700
SN  - 1520-6882
ER  -