Increased coverage in the transmembrane domain with activated-ion electron capture dissociation for top-down Fourier-transform mass spectrometry of integral membrane proteins.
The c-subunit of ATP synthase (AtpH) is an 8 kD integral membrane protein with two transmembrane domains; we set out to demonstrate it amenable to top-down electrospray-ionization Fourier-transform mass spectrometry (FT-MS) using both collision activated and electron capture dissociation (CAD/ECD). Thermal activation concomitant with electron delivery was necessary for efficient ECD (activated-ion ECD; aiECD), yielding complementary information and greater sequence coverage in the transmembrane domains in comparison with CAD.
Zabrouskov
Vlad
V
Thermo Fisher, San Jose, California, California 95134, USA.
Whitelegge
Julian P
JP
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry; D004583:Electrons; D007477:Ions; D008565:Membrane Proteins / Q000032:analysis*; D025261:Mitochondrial Proton-Translocating ATPases / Q000032:analysis*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D040901:Proteomics / Q000379:methods*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D007477:Ions; D008565:Membrane Proteins; C103607:mitochondrial ATPase subunit c; D025261:Mitochondrial Proton-Translocating ATPases