Peptide and protein characterization by high-rate electron capture dissociation Fourier transform ion cyclotron resonance mass spectrometry.
The analytical utility of the electron capture dissociation (ECD) technique, developed by McLafferty and co-workers, has substantially improved peptide and protein characterization using Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS). The limitations of the first ECD implementations on commercial instruments were eliminated by the employment of low-energy electron-injection systems based on indirectly heated dispenser cathodes. In particular, the ECD rate and reliability were greatly increased, enabling the combination of ECD/FTICR-MS with on-line liquid separation techniques. Further technique development allowed the combination of two rapid fragmentation techniques, high-rate ECD and infrared multiphoton dissociation (IRMPD), in a single experimental configuration. Simultaneous and consecutive irradiations of trapped ions with electrons and photons extended the possibilities for ion activation/dissociation and led to improved peptide and protein characterization. The application of high-rate ECD/FTICR-MS has demonstrated its power and unique capabilities in top-down sequencing of peptides and proteins, including characterization of post-translational modifications, improved sequencing of peptides with multiple disulfide bridges and secondary fragmentation (w-ion formation). Analysis of peptide mixtures has been accomplished using high-rate ECD in bottom-up mass spectrometry based on mixture separation by liquid chromatography and capillary electrophoresis. This paper summarizes the current impact of high-rate ECD/FTICR-MS for top-down and bottom-up mass spectrometry of peptides and proteins.
Division of Ion Physics, Angström Laboratory, Uppsala University, Box 534, SE-751 21, Uppsala, Sweden. youri.tsybin@angstrom.uu.se
Youri O
YO
Tsybin
Margareta
M
Ramström
Matthias
M
Witt
Gökhan
G
Baykut
Per
P
Håkansson
D000818:Animals; D002417:Cattle; D017357:Cyclotrons; D006801:Humans; D010455:Peptides / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D020539:Sequence Analysis, Protein; D012709:Serum Albumin / Q000032:analysis; D017550:Spectroscopy, Fourier Transform Infrared / Q000295:instrumentation / Q000379:methods*; D025801:Ubiquitin / Q000032:analysis
D010455:Peptides; D011506:Proteins; D012709:Serum Albumin; D025801:Ubiquitin