Mass spectrometry of membrane transporters reveals subunit stoichiometry and interactions.
We describe a general mass spectrometry approach to determine subunit stoichiometry and lipid binding in intact membrane protein complexes. By exploring conditions for preserving interactions during transmission into the gas phase and for optimally stripping away detergent, by subjecting the complex to multiple collisions, we released the intact complex largely devoid of detergent. This enabled us to characterize both subunit stoichiometry and lipid binding in 4 membrane protein complexes.
Barrera
Nelson P
NP
Department of Chemistry, University of Cambridge, Cambridge, UK.
Isaacson
Shoshanna C
SC
Zhou
Min
M
Bavro
Vassiliy N
VN
Welch
Alex
A
Schaedler
Theresia A
TA
Seeger
Markus A
MA
Miguel
Ricardo Núñez
RN
Korkhov
Vladimir M
VM
van Veen
Hendrik W
HW
Venter
Henrietta
H
Walmsley
Adrian R
AR
Tate
Christopher G
CG
Robinson
Carol V
CV
D026901:Membrane Transport Proteins / Q000737:chemistry*; D046912:Multiprotein Complexes / Q000737:chemistry*; D025941:Protein Interaction Mapping; D021122:Protein Subunits / Q000737:chemistry*; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D026901:Membrane Transport Proteins; D046912:Multiprotein Complexes; D021122:Protein Subunits