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

doi:10.1007/s13361-017-1799-4

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

Abstract

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.

Authors:

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)

Publication Date:: 2017-10-02

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

Sponsoring Org.:: USDOE

OSTI Identifier:: 1511586

Grant/Contract Number:: FC03-02ER63421

Resource Type:: Accepted Manuscript

Journal Name:: Journal of the American Society for Mass Spectrometry

Additional Journal Information:: Journal Volume: 29; Journal Issue: 1; Journal ID: ISSN 1044-0305

Publisher:: American Society for Mass Spectrometry

Country of Publication:: United States

Language:: English

Subject:: 59 BASIC BIOLOGICAL SCIENCES; 37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY; Native-mass spectrometry; Membrane protein; Fourier transform ion cyclotron resonance

Citation Formats


                    Lippens, Jennifer L., Nshanian, Michael, Spahr, Chris, Egea, Pascal F., Loo, Joseph A., and Campuzano, Iain D.  G. Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes.  United States: N. p., 2017. 
        Web.  doi:10.1007/s13361-017-1799-4.

Copy to clipboard


                    Lippens, Jennifer L., Nshanian, Michael, Spahr, Chris, Egea, Pascal F., Loo, Joseph A., & Campuzano, Iain D.  G. Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes.  United States.  doi:10.1007/s13361-017-1799-4.

Copy to clipboard


                    Lippens, Jennifer L., Nshanian, Michael, Spahr, Chris, Egea, Pascal F., Loo, Joseph A., and Campuzano, Iain D.  G. Mon .  
        "Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes".  United States.  doi:10.1007/s13361-017-1799-4.  https://www.osti.gov/servlets/purl/1511586.

Copy to clipboard


                    
@article{osti_1511586,

  title        = {Fourier Transform-Ion Cyclotron Resonance Mass Spectrometry as a Platform for Characterizing Multimeric Membrane Protein Complexes},

  author       = {Lippens, Jennifer L. and Nshanian, Michael and Spahr, Chris and Egea, Pascal F. and Loo, Joseph A. and Campuzano, Iain D.  G.},

  abstractNote = {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.},

  doi          = {10.1007/s13361-017-1799-4},

  journal      = {Journal of the American Society for Mass Spectrometry},

  number       = 1,

  volume       = 29,

  place        = {United States},

  year         = {2017},

  month        = {10}

}

Copy to clipboard

Journal Article:

Free Publicly Available Full Text

Accepted Manuscript (DOE)

Publisher's Version of Record

DOI: 10.1007/s13361-017-1799-4

Other availability

Search WorldCat to find libraries that may hold this journal

Save / Share:

Export Metadata

Save to My Library

Works referenced in this record:

Self-Assembly of Discoidal Phospholipid Bilayer Nanoparticles with Membrane Scaffold Proteins
journal, August 2002

Bayburt, Timothy H.; Grinkova, Yelena V.; Sligar, Stephen G.
Nano Letters, Vol. 2, Issue 8, p. 853-856
DOI: 10.1021/nl025623k

Similar Records in DOE PAGES and OSTI.GOV collections:

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

Journal Article Lippens, Jennifer L. ; Nshanian, Michael ; Spahr, Chris ; ... - Journal of the American Society for Mass Spectrometry

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 Q-ToF, 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 degreesmore »« less
DOI: 10.1007/S13361-017-1799-4
Surface-Induced Dissociation of Protein Complexes in a Hybrid Fourier Transform Ion Cyclotron Resonance Mass Spectrometer

Journal Article Yan, Jing ; Zhou, Mowei ; Gilbert, Joshua D. ; ... - Analytical Chemistry

Mass spectrometry continues to develop as a valuable tool in the analysis of proteins and protein complexes. In protein complex mass spectrometry studies, surface-induced dissociation (SID) has been successfully applied in quadrupole time-of-flight (Q-TOF) instruments. SID provides structural information on noncovalent protein complexes that is complementary to other techniques. However, the mass resolution of Q-TOF instruments can limit the information that can be obtained for protein complexes by SID. Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) provides ultrahigh resolution and ultrahigh mass accuracy measurements. Here in this study, an SID device was designed and successfully installed in amore »« less
Cited by 2
DOI: 10.1021/acs.analchem.6b03986

Full Text Available
Integration of Electrokinetic-Based Multidimensional Separations/Concentration Platform with electrospray ionization-Fourier transform ion cyclotron resonance-mass spectrometry for Proteome Analysis of Shewanella Oneidensis

Journal Article Mohan, Deepa ; Pasa-Tolic, Ljiljana ; Masselon, Christophe D. ; ... - Analytical Chemistry

No abstract prepared.
DOI: 10.1021/ac0342572
Efficacy of bacterial bioremediation: Demonstration of complete incorporation of hydrocarbons into membrane phospholipids from Rhodococcus hydrocarbon degrading bacteria by electrospray ionization Fourier transform ion cyclotron resonance mass spectrometry

Journal Article Rodgers, R.P. ; Blumer, E.N. ; Emmett, M.R. ; ... - Environmental Science and Technology

The authors present a method and example to establish complete incorporation of hydrocarbons into membrane phospholipids of putatively bioremediative bacteria. Bacteria are grown on minimal media containing a specified carbon source, either natural abundance or enriched. After extraction (but no other prior separation) of the membrane lipids, electrospray ionization yields a negative-ion FT-ICR mass spectrum containing prominent phospholipid parent ions. If {sup 13}C-enriched hydrocarbon incorporation is complete, then the mass of the parent ion will increase by n Da, in which n is the number of its constituent carbon atoms; moreover, the {sup 13}C isotopic distribution pattern will be reversed.more »« less
DOI: 10.1021/es990889n
Isotope-labeled cross-linkers and fourier transform ion cyclotron resonance mass spectrometry for structural analysis of a protein/peptide complex

Journal Article Ihling, Christian ; Schmidt, Andreas ; Kalkhof, Stefan ; ... - Journal of the American Society for Mass Spectrometry

For structural studies of proteins and their complexes, chemical cross-linking combined with mass spectrometry presents a promising strategy to obtain structural data of protein interfaces from low quantities of proteins within a short time. We explore the use of isotope-labeled cross-linkers in combination with Fourier transform ion cyclotron resonance (FTICR) mass spectrometry for a more efficient identification of cross-linker containing species. For our studies, we chose the calcium-independent complex between calmodulin and a 25-amino acid peptide from the C-terminal region of adenylyl cyclase 8 containing an “IQ-like motif.” Cross-linking reactions between calmodulin and the peptide were performed in the absencemore »« less
DOI: 10.1016/J.JASMS.2006.04.020

Similar Records

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

Abstract

Citation Formats

Self-Assembly of Discoidal Phospholipid Bilayer Nanoparticles with Membrane Scaffold Proteins journal, August 2002

Self-Assembly of Discoidal Phospholipid Bilayer Nanoparticles with Membrane Scaffold Proteins
journal, August 2002