Isotopic Resolution of Protein Complexes up to 466 kDa Using Individual Ion Mass Spectrometry

John P. McGee; Rafael D. Melani; Ping F. Yip; Michael W. Senko; Philip D. Compton; Jared O. Kafader; Neil L. Kelleher

doi:10.1021/acs.analchem.0c03282

Isotopic Resolution of Protein Complexes up to 466 kDa Using Individual Ion Mass Spectrometry

John P. McGee, Rafael D. Melani, Ping F. Yip, Michael W. Senko, Philip D. Compton, Jared O. Kafader, Neil L. Kelleher^*

^*Corresponding author for this work

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

42 Scopus citations

Abstract

Native mass spectrometry involves transferring large biomolecular complexes into the gas phase, enabling the characterization of their composition and stoichiometry. However, the overlap in distributions created by residual solvation, ionic adducts, and post-translational modifications creates a high degree of complexity that typically goes unresolved at masses above ∼150 kDa. Therefore, native mass spectrometry would greatly benefit from higher resolution approaches for intact proteins and their complexes. By recording mass spectra of individual ions via charge detection mass spectrometry, we report isotopic resolution for pyruvate kinase (232 kDa) and β-galactosidase (466 kDa), extending the limits of isotopic resolution for high mass and high m/z by >2.5-fold and >1.6-fold, respectively.

Original language	English (US)
Journal	Analytical Chemistry
Volume	93
Issue number	5
DOIs	https://doi.org/10.1021/acs.analchem.0c03282
State	Published - Feb 9 2021

Funding

This work was completed with support from the Intensifying Innovation (I) program at Thermo Fisher Scientific and was carried out in collaboration with the National Resource for Translational and Developmental Proteomics under Grant P41 GM108569 from the National Institute of General Medical Sciences, National Institutes of Health, RF1 AG063903 from the National Institute on Aging, and an instrumentation award (S10 OD025194) from the NIH Office of the Director. Further support from an F31 Fellowship to J.P.M. (F31 AG069456) and support from the National Institute on Drug Abuse (P30 DA018310) is also acknowledged. 2

ASJC Scopus subject areas

Analytical Chemistry

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abstract = "Native mass spectrometry involves transferring large biomolecular complexes into the gas phase, enabling the characterization of their composition and stoichiometry. However, the overlap in distributions created by residual solvation, ionic adducts, and post-translational modifications creates a high degree of complexity that typically goes unresolved at masses above ∼150 kDa. Therefore, native mass spectrometry would greatly benefit from higher resolution approaches for intact proteins and their complexes. By recording mass spectra of individual ions via charge detection mass spectrometry, we report isotopic resolution for pyruvate kinase (232 kDa) and β-galactosidase (466 kDa), extending the limits of isotopic resolution for high mass and high m/z by >2.5-fold and >1.6-fold, respectively.",

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AU - McGee, John P.

AU - Melani, Rafael D.

AU - Yip, Ping F.

AU - Senko, Michael W.

AU - Compton, Philip D.

AU - Kafader, Jared O.

AU - Kelleher, Neil L.

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Isotopic Resolution of Protein Complexes up to 466 kDa Using Individual Ion Mass Spectrometry

Abstract

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