Cited by 71Mass Spectrometry Techniques and ApplicationsAdvanced Electron Microscopy Techniques and ApplicationsAdvanced Proteomics Techniques and Applications

Mass-Selective Soft-Landing of Protein Assemblies with Controlled Landing Energies

Mikhailov, Victor A., Mize, Todd H., Benesch, Justin L. P., Robinson, Carol V.·2014·Analytical Chemistry

Abstract

Selection and soft-landing of bionanoparticles in vacuum is potentially a preparative approach to separate heterogeneous mixtures for high-resolution structural study or to deposit homogeneous materials for nanotechnological applications. Soft-landing of intact protein assemblies however remains challenging, due to the difficulties of manipulating these heavy species in mass-selective devices and retaining their structure during the experiment. We have developed a tandem mass spectrometer with the capability for controlled ion soft-landing and ex situ visualization of the soft-landed particles by means of transmission electron microscopy. The deposition conditions can be controlled by adjusting the kinetic energies of the ions by applying accelerating or decelerating voltages to a set of ion-steering optics. To validate this approach, we have examined two cage-like protein complexes, GroEL and ferritin, and studied the effect of soft-landing conditions on the method's throughput and the preservation of protein structure. Separation, based on mass-to-charge ratio, of holo- and apo-ferritin complexes after electrospray ionization enabled us to soft-land independently the separated complexes on a grid suitable for downstream transmission electron microscopy analysis. Following negative staining, images of the soft-landed complexes reveal that their structural integrity is largely conserved, with the characteristic central cavity of apoferritin, and iron core of holoferritin, surviving the phase transition from liquid to gas, soft-landing, and dehydration in vacuum.

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Crossref · 10.1021/ac5018327

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Authors
Mikhailov, V. A., Mize, T. H., Benesch, J. L. P., & Robinson, C. V.
Affiliations
University of Oxford (GB)
Corresponding author
  • Benesch, Justin L. P.
  • Robinson, Carol V.
Venue
Analytical Chemistry
Published
2014-07-31
Volume
86
Issue
16
Pages
8321-8328
ISSN
  • 0003-2700 (ISSN-L)
  • 1520-6882
OpenAlex
W2068602940
Cited by
71
References
53
Citations by year
2026: 3 · 2025: 5 · 2024: 6 · 2023: 10 · 2022: 13 · 2021: 7 · 2020: 5 · 2019: 4 · 2018: 3 · 2017: 4 · 2016: 4 · 2015: 6 · 2014: 1
FWCI
2.14
Type
journal article
Language
en
Open access
closed
Locations
  • Analytical Chemistry (publishedVersion)
  • PubMed (publishedVersion)
  • University of Birmingham Research Portal (University of Birmingham) (submittedVersion)
Funders
  • Medical Research Council
  • Engineering and Physical Sciences Research Council
  • European Union PROSPECTS Network
Topics
  • Physical Sciences › Chemistry › Spectroscopy › Mass Spectrometry Techniques and Applications
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Structural Biology › Advanced Electron Microscopy Techniques and Applications
  • Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
Keywords
tandem mass spectrometry, protein assembly, bionanoparticles, GroEL, ferritin, electrospray ionization, protein structural integrity, vacuum deposition
MeSH terms
Animals, Equipment Design, Escherichia coli, Ferritins, Horses, Ions, Kinetics, Vacuum, Chaperonin 60, Escherichia coli Proteins, Tandem Mass Spectrometry, Nanoparticles

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