Easy amino acid sequencing of sulfonated peptides using post-source decay on a matrix-assisted laser desorption/ionization time-of-flight mass spectrometer equipped with a variable voltage reflector.
Tryptic peptides were labeled with sulfonic acid groups at the N-termini using an improved chemistry. The derivatization was performed in common aqueous buffers on peptides adsorbed onto a ZipTip trade mark C(18), thus allowing simultaneous desalting/concentration of the sample. When only Arg-terminating peptides were considered, the procedure from adsorption onto the ZipTip until analysis by MALDI-PSD took about 10 min and several samples could be worked on in parallel. The resulting improved post-source decay (PSD) fragmentation produced spectra containing only y-ions. PSD amino acid sequencing of underivatized and derivatized synthetic peptides was compared. From the sequence information obtained from derivatized peptides isolated by ion selection from tryptic in-gel digests, a protein was correctly identified which was difficult to analyze from an unclear peptide mass fingerprint analysis. The method was also applied to the identification and localization of phosphorylated Ser and Tyr residues in native and synthetic peptides.
Hellman
Ulf
U
Ludwig Institute for Cancer Research, Box 595, SE-751 24 Uppsala, Sweden. ulf.hellman@licr.uu.se
Bhikhabhai
Rama
R
D006146:Guanidines / Q000737:chemistry; D006868:Hydrolysis; D007202:Indicators and Reagents; D008239:Lysine / Q000737:chemistry; D010455:Peptides / Q000032:analysis*; D010748:Phosphopeptides / Q000032:analysis; D017421:Sequence Analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013451:Sulfonic Acids / Q000032:analysis; D014357:Trypsin
D006146:Guanidines; D007202:Indicators and Reagents; D010455:Peptides; D010748:Phosphopeptides; D013451:Sulfonic Acids; D014357:Trypsin; D008239:Lysine