Negative ions enhance survival of membrane protein complexes
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...
| Главные авторы: | , , , , |
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| Формат: | Journal article |
| Язык: | English |
| Опубликовано: |
Springer Verlag
2016
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| _version_ | 1826310714269106176 |
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| author | Liko, I Allison, T Hopper, J Benesch, J Robinson, C |
| author_facet | Liko, I Allison, T Hopper, J Benesch, J Robinson, C |
| author_sort | Liko, I |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T07:56:03Z |
| format | Journal article |
| id | oxford-uuid:75a878dc-ee9d-45c0-b3da-e91ea5a849b9 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T07:56:03Z |
| publishDate | 2016 |
| publisher | Springer Verlag |
| record_format | dspace |
| spelling | oxford-uuid:75a878dc-ee9d-45c0-b3da-e91ea5a849b92023-08-21T10:42:49ZNegative ions enhance survival of membrane protein complexesJournal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:75a878dc-ee9d-45c0-b3da-e91ea5a849b9EnglishSymplectic Elements at OxfordSpringer Verlag2016Liko, IAllison, THopper, JBenesch, JRobinson, CMembrane 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. |
| spellingShingle | Liko, I Allison, T Hopper, J Benesch, J Robinson, C Negative ions enhance survival of membrane protein complexes |
| title | Negative ions enhance survival of membrane protein complexes |
| title_full | Negative ions enhance survival of membrane protein complexes |
| title_fullStr | Negative ions enhance survival of membrane protein complexes |
| title_full_unstemmed | Negative ions enhance survival of membrane protein complexes |
| title_short | Negative ions enhance survival of membrane protein complexes |
| title_sort | negative ions enhance survival of membrane protein complexes |
| work_keys_str_mv | AT likoi negativeionsenhancesurvivalofmembraneproteincomplexes AT allisont negativeionsenhancesurvivalofmembraneproteincomplexes AT hopperj negativeionsenhancesurvivalofmembraneproteincomplexes AT beneschj negativeionsenhancesurvivalofmembraneproteincomplexes AT robinsonc negativeionsenhancesurvivalofmembraneproteincomplexes |