Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes

Lippens, Jennifer L.; Nshanian, Michael; Spahr, Chris; Egea, Pascal F.; Loo, Joseph A.; Campuzano, Iain D.  G.

doi:10.1007/s13361-017-1799-4

Title: Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes

Journal Article · 02 October 2017 · Journal of the American Society for Mass Spectrometry

DOI: https://doi.org/10.1007/s13361-017-1799-4 · OSTI ID:1511586

Lippens, Jennifer L. ^[1]; Nshanian, Michael ^[2]; Spahr, Chris ^[1]; Egea, Pascal F. ^[2]; Loo, Joseph A. ^[2]; Campuzano, Iain D. G. ^[1]

Discovery Attribute Sciences, Amgen, Thousand Oaks, CA (United States)
Univ. of California, Los Angeles, CA (United States)

We report that membrane protein characterization is consistently hampered by challenges with expression, purification and solubilization. Among several biophysical techniques employed for their characterization, native-mass spectrometry (MS) has emerged as a powerful tool for the analysis of membrane proteins and complexes. Here, two MS platforms, the FT-ICR and QToF, have been explored to analyze the homotetrameric water channel protein, AquaporinZ (AqpZ) under non-denaturing conditions. This 97 kDa membrane protein complex can be readily liberated from the octylglucoside (OG) detergent micelle under a range of instrument conditions on both MS platforms. Increasing the applied collision energy of the FT-ICR collision cell yielded varying degrees of tetramer (97 kDa) liberation from the OG micelles, as well as dissociation into the trimeric (72 kDa) and monomeric (24 kDa) substituents. Tandem-MS on the Q-ToF yielded higher intensity tetramer signal and, depending on the m/z region selected, the observed monomer signal varied in intensity. Precursor ion selection of an m/z range above the expected protein signal distribution, followed by mild collisional activation is able to efficiently liberate AqpZ with a high S/N ratio. Lastly, the tetrameric charge state distribution obtained on both instruments demonstrated superpositioning of multiple proteoforms due to varying degrees of N-terminal formylation.

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Research Organization:: Univ. of California, Los Angeles, CA (United States)

Sponsoring Organization:: USDOE

Grant/Contract Number:: FC03-02ER63421

OSTI ID:: 1511586

Journal Information:: Journal of the American Society for Mass Spectrometry, Vol. 29, Issue 1; ISSN 1044-0305

Publisher:: American Society for Mass SpectrometryCopyright Statement

Country of Publication:: United States

Language:: English

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Cited by: 26 works

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Native-mass spectrometry
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