Charge Reduction Stabilizes Intact Membrane Protein Complexes for Mass Spectrometry

Overview

Journal J Am Chem Soc

Publisher American Chemical Society

Specialty Chemistry

Date 2014 Nov 18

PMID 25402655

Citations 44

Authors

Shahid Mehmood

Julien Marcoux

Jonathan T S Hopper

Timothy M Allison

Idlir Liko

Antoni J Borysik

Carol V Robinson

Affiliations

Soon will be listed here.

Abstract

The study of intact soluble protein assemblies by means of mass spectrometry is providing invaluable contributions to structural biology and biochemistry. A recent breakthrough has enabled similar study of membrane protein complexes, following their release from detergent micelles in the gas phase. Careful optimization of mass spectrometry conditions, particularly with respect to energy regimes, is essential for maintaining compact folded states as detergent is removed. However, many of the saccharide detergents widely employed in structural biology can cause unfolding of membrane proteins in the gas phase. Here, we investigate the potential of charge reduction by introducing three membrane protein complexes from saccharide detergents and show how reducing their overall charge enables generation of compact states, as evidenced by ion mobility mass spectrometry. We find that charge reduction stabilizes the oligomeric state and enhances the stability of lipid-bound complexes. This finding is significant since maintaining native-like membrane proteins enables ligand binding to be assessed from a range of detergents that retain solubility while protecting the overall fold.

Citing Articles

Structural Analysis of the 20S Proteasome Using Native Mass Spectrometry and Ultraviolet Photodissociation.

Walker J, Gautam A, Matouschek A, Brodbelt J J Proteome Res. 2024; 23(12):5438-5448.

PMID: 39475212 PMC: 11660092. DOI: 10.1021/acs.jproteome.4c00568.

Native Mass Spectrometry of Membrane Protein-Lipid Interactions in Different Detergent Environments.

Kumar S, Stover L, Wang L, Bahramimoghaddam H, Zhou M, Russell D Anal Chem. 2024; 96(42):16768-16776.

PMID: 39394983 PMC: 11503522. DOI: 10.1021/acs.analchem.4c03312.

To 200,000 / and Beyond: Native Electron Capture Charge Reduction Mass Spectrometry Deconvolves Heterogeneous Signals in Large Biopharmaceutical Analytes.

Le Huray K, Worner T, Moreira T, Dembek M, Reinhardt-Szyba M, Devine P ACS Cent Sci. 2024; 10(8):1548-1561.

PMID: 39220705 PMC: 11363327. DOI: 10.1021/acscentsci.4c00462.

Native mass spectrometry of membrane protein-lipid interactions in different detergent environments.

Kumar S, Stover L, Wang L, Bahramimoghaddam H, Zhou M, Russell D bioRxiv. 2024; .

PMID: 38979331 PMC: 11230385. DOI: 10.1101/2024.06.27.601044.

Evaluation of a Commercial TIMS-Q-TOF Platform for Native Mass Spectrometry.

Panczyk E, Lin Y, Harvey S, Snyder D, Liu F, Ridgeway M J Am Soc Mass Spectrom. 2024; 35(7):1394-1402.

PMID: 38905538 PMC: 11651300. DOI: 10.1021/jasms.3c00320.

References

Benesch J, Aquilina J, Ruotolo B, Sobott F, Robinson C . Tandem mass spectrometry reveals the quaternary organization of macromolecular assemblies. Chem Biol. 2006; 13(6):597-605. DOI: 10.1016/j.chembiol.2006.04.006. View

Marcoux J, Politis A, Rinehart D, Marshall D, Wallace M, Tamm L . Mass spectrometry defines the C-terminal dimerization domain and enables modeling of the structure of full-length OmpA. Structure. 2014; 22(5):781-90. PMC: 4147082. DOI: 10.1016/j.str.2014.03.004. View

Borysik A, Hewitt D, Robinson C . Detergent release prolongs the lifetime of native-like membrane protein conformations in the gas-phase. J Am Chem Soc. 2013; 135(16):6078-83. DOI: 10.1021/ja401736v. View

Marcoux J, Wang S, Politis A, Reading E, Ma J, Biggin P . Mass spectrometry reveals synergistic effects of nucleotides, lipids, and drugs binding to a multidrug resistance efflux pump. Proc Natl Acad Sci U S A. 2013; 110(24):9704-9. PMC: 3683783. DOI: 10.1073/pnas.1303888110. View

Parker J, Newstead S . Current trends in α-helical membrane protein crystallization: an update. Protein Sci. 2012; 21(9):1358-65. PMC: 3631364. DOI: 10.1002/pro.2122. View

Catalina M, van den Heuvel R, van Duijn E, Heck A . Decharging of globular proteins and protein complexes in electrospray. Chemistry. 2004; 11(3):960-8. DOI: 10.1002/chem.200400395. View

Ruotolo B, Hyung S, Robinson P, Giles K, Bateman R, Robinson C . Ion mobility-mass spectrometry reveals long-lived, unfolded intermediates in the dissociation of protein complexes. Angew Chem Int Ed Engl. 2007; 46(42):8001-4. DOI: 10.1002/anie.200702161. View

Seddon A, Curnow P, Booth P . Membrane proteins, lipids and detergents: not just a soap opera. Biochim Biophys Acta. 2004; 1666(1-2):105-17. DOI: 10.1016/j.bbamem.2004.04.011. View

Pagel K, Hyung S, Ruotolo B, Robinson C . Alternate dissociation pathways identified in charge-reduced protein complex ions. Anal Chem. 2010; 82(12):5363-72. DOI: 10.1021/ac101121r. View

10.

Morgner N, Montenegro F, Barrera N, Robinson C . Mass spectrometry--from peripheral proteins to membrane motors. J Mol Biol. 2012; 423(1):1-13. PMC: 4058634. DOI: 10.1016/j.jmb.2012.06.033. View

11.

Hall Z, Politis A, Bush M, Smith L, Robinson C . Charge-state dependent compaction and dissociation of protein complexes: insights from ion mobility and molecular dynamics. J Am Chem Soc. 2012; 134(7):3429-38. DOI: 10.1021/ja2096859. View

12.

Laganowsky A, Reading E, Allison T, Ulmschneider M, Degiacomi M, Baldwin A . Membrane proteins bind lipids selectively to modulate their structure and function. Nature. 2014; 510(7503):172-175. PMC: 4087533. DOI: 10.1038/nature13419. View

13.

Ward A, Szewczyk P, Grimard V, Lee C, Martinez L, Doshi R . Structures of P-glycoprotein reveal its conformational flexibility and an epitope on the nucleotide-binding domain. Proc Natl Acad Sci U S A. 2013; 110(33):13386-91. PMC: 3746859. DOI: 10.1073/pnas.1309275110. View

14.

Hopper J, Sokratous K, Oldham N . Charge state and adduct reduction in electrospray ionization-mass spectrometry using solvent vapor exposure. Anal Biochem. 2011; 421(2):788-90. DOI: 10.1016/j.ab.2011.10.034. View

15.

Scalf M, Westphall M, Krause J, Kaufman S, Smith L . Controlling charge states of large ions. Science. 1999; 283(5399):194-7. DOI: 10.1126/science.283.5399.194. View

16.

Barrera N, Di Bartolo N, Booth P, Robinson C . Micelles protect membrane complexes from solution to vacuum. Science. 2008; 321(5886):243-6. DOI: 10.1126/science.1159292. View

17.

Housden N, Wojdyla J, Korczynska J, Grishkovskaya I, Kirkpatrick N, Brzozowski A . Directed epitope delivery across the Escherichia coli outer membrane through the porin OmpF. Proc Natl Acad Sci U S A. 2010; 107(50):21412-7. PMC: 3003033. DOI: 10.1073/pnas.1010780107. View

18.

Testa L, Brocca S, Grandori R . Charge-surface correlation in electrospray ionization of folded and unfolded proteins. Anal Chem. 2011; 83(17):6459-63. DOI: 10.1021/ac201740z. View

19.

Hopper J, Oldham N . Alkali metal cation-induced destabilization of gas-phase protein-ligand complexes: consequences and prevention. Anal Chem. 2011; 83(19):7472-9. DOI: 10.1021/ac201686f. View

20.

Sun D, Rosokha S, Kochi J . Reversible interchange of charge-transfer versus electron-transfer states in organic electron transfer via cross-exchanges between diamagnetic (donor/acceptor) dyads. J Phys Chem B. 2007; 111(24):6655-66. DOI: 10.1021/jp068994o. View