An enhanced method for peptides sequencing by N-terminal derivatization and MS.
An improved method for peptide sequencing based on acetylation/deuteroacetylation in conjunction with ESI MS is introduced. Derivatization with a 1:1 mixture of acetic anhydride and deuterated acetic anhydride incorporates a stable isotope label into the analyzed molecule. This approach has been initially applied to FAB. Using MS/MS, the technique provides a fast, highly sensitive and reliable determination of the primary structure of unknown peptides. This procedure labels N-terminal fragments formed during MS/MS analysis, resulting in a simplification and faster interpretation of the spectra. The performance of the method has been tested with several synthetic peptides and applied to an efficient sequencing of the peptide map, using a nano-scale LC coupled on-line to a tandem mass spectrometer.
Noga
Marek J
MJ
Department of Neurobiochemistry, Faculty of Chemistry and Regional Laboratory, Jagiellonian University, Krakow, Poland.
Lewandowski
Jakub J
JJ
Suder
Piotr
P
Silberring
Jerzy
J
D000107:Acetylation; D045304:Cytochromes c / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods; D009211:Myoglobin / Q000737:chemistry; D010446:Peptide Fragments / Q000737:chemistry; D010449:Peptide Mapping / Q000379:methods; D010455:Peptides / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D014357:Trypsin / Q000737:chemistry
D009211:Myoglobin; D010446:Peptide Fragments; D010455:Peptides; D045304:Cytochromes c; D014357:Trypsin