Title
Top-down mass spectrometry of intact membrane protein complexes reveals oligomeric state and sequence information in a single experiment Top-down mass spectrometry of intact membrane protein complexes reveals oligomeric state and sequence information in a single experiment
Author
Faculty/Department
Faculty of Sciences. Chemistry
Publication type
article
Publication
Cambridge ,
Subject
Chemistry
Biology
Source (journal)
Protein science. - Cambridge
Volume/pages
24(2015) :8 , p. 1292-1300
ISSN
0961-8368
ISI
000358176100012
Carrier
E
Target language
English (eng)
Full text (Publishers DOI)
Affiliation
University of Antwerp
Abstract
Here we study the intact stoichiometry and top-down fragmentation behavior of three integral membrane proteins which were natively reconstituted into detergent micelles: the mechano-sensitive ion channel of large conductance (MscL), the Kirbac potassium channel and the p7 viroporin from the hepatitis C virus. By releasing the proteins under nondenaturing conditions inside the mass spectrometer, we obtained their oligomeric sizes. Increasing the ion activation (collision energy) causes unfolding and subsequent ejection of a highly charged monomer from the membrane protein complexes. Further increase of the ion activation then causes collision-induced dissociation (CID) of the ejected monomers, with fragments observed which were predominantly found to stem from membrane-embedded regions. These experiments show how in a single experiment, we can probe the relation between higher-order structure and protein sequence, by combining the native MS data with fragmentation obtained from top-down MS.
Full text (open access)
https://repository.uantwerpen.be/docman/irua/904e05/10467.pdf
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