A simple and effective method was developed for peptide sequencing and protein identification through the determination of its N-terminal residue. The method of N-terminal carbamidomethylation with iodoacetamide could specifically and remarkably enhance the intensity of a1 ions in the tandem mass spectra of the peptide derivatives without significantly altering their fragmentation pattern, thus allowing determination of their N-terminal residues. The effectiveness and specificity of the method was demonstrated by confirming and extending sequence interpretation of several model peptides and proteins. The developed method was then applied in the LC-MS/MS analysis of the tryptic digests of myoglobin and a whole protein extract from rat heart tissues. The results from database searches were well validated with the enhancement of a1 ions in tandem mass spectra and the specificity of protein identification was obtained when the information of N-terminal residues was included in the database search.
|Number of pages||10|
|Journal||Journal of the American Society for Mass Spectrometry|
|Publication status||Published - Jun 2009|
Scopus Subject Areas
- Structural Biology