In-trap cleanup of proteins from electrospray ionization using soft sustained off-resonance irradiation with fourier transform ion cyclotron resonance mass spectrometry.
Electrospray ionization Fourier transform ion cyclotron resonance (FTICR) mass spectrometry is capable of producing high mass resolving power and improved mass accuracy for large proteins and noncovalent complexes when coupled with collisionally induced dissociation (CID) of noncovalent adducts and consequent minimization of ion charge density in the ICR trap during measurements. This work demonstrates the application of in-trap cleanup to several biologically relevant systems, including carbonic anhydrase, 4-oxalocrotonate tautomerase (4OT) analogue, and SecB, a chaperone from Escherichia coli. In-trap cleanup yields improved mass measurements for these systems and is expected to further enable measurements for even more complex systems where adduction levels have precluded study of intact complexes.
Tolić
L P
LP
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, Washington 99352, USA.
Bruce
J E
JE
Lei
Q P
QP
Anderson
G A
GA
Smith
R D
RD
D001426:Bacterial Proteins / Q000737:chemistry; D002256:Carbonic Anhydrases / Q000737:chemistry; D017357:Cyclotrons; D004926:Escherichia coli; D007535:Isomerases / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D018832:Molecular Chaperones / Q000737:chemistry; D011506:Proteins; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D001426:Bacterial Proteins; D018832:Molecular Chaperones; D011506:Proteins; C057842:SecB protein, Bacteria; D002256:Carbonic Anhydrases; C021918:4-oxalocrotonate tautomerase; D007535:Isomerases