Journal
JOURNAL OF THE AMERICAN SOCIETY FOR MASS SPECTROMETRY
Volume 27, Issue 6, Pages 1099-1104Publisher
SPRINGER
DOI: 10.1007/s13361-016-1381-5
Keywords
Membrane proteins; Negative polarity; Structural biology
Funding
- ERC [26851]
- Royal Society Research Professorship
- Medical Research Council [98101]
- Engineering and Physical Sciences Research Council [EP/J01835X/1] Funding Source: researchfish
- Medical Research Council [G1000819] Funding Source: researchfish
- EPSRC [EP/J01835X/1] Funding Source: UKRI
- MRC [G1000819] Funding Source: UKRI
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Membrane protein complexes are commonly introduced to the mass spectrometer solubilized in detergent micelles. The collisional activation used to remove the detergent, however, often causes protein unfolding and dissociation. As in the case for soluble proteins, electrospray in the positive ion mode is most commonly used for the study of membrane proteins. Here we show several distinct advantages of employing the negative ion mode. Negative polarity can yield lower average charge states for membrane proteins solubilized in saccharide detergents, with enhanced peak resolution and reduced adduct formation. Most importantly, we demonstrate that negative ion mode electrospray ionization (ESI) minimizes subunit dissociation in the gas phase, allowing access to biologically relevant oligomeric states. Together, these properties mean that intact membrane protein ions can be generated in a greater range of solubilizing detergents. The formation of negative ions, therefore, greatly expands the possibilities of using mass spectrometry on this intractable class of protein.
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