Capture of peptides with N-terminal serine and threonine: a sequence-specific chemical method for Peptide mixture simplification.
The objective of this study was to evaluate a sequence-specific chemistry for the ability to specifically capture peptides that contain N-terminal serine or threonine residues from mixtures. The first step is the oxidation of the 1,2-amino alcohol structure -CH(NH(2))CH(OH)- of peptides containing N-terminal serine or threonine with periodate. The newly formed aldehyde reacts with a labeling reagent containing a hydrazide, RCONHNH(2), to form a hydrazone-peptide conjugate, RCONHN=CH-peptide. Biotin-labeled conjugates can then be isolated by affinity purification with streptavidin. The method described in this report can be useful in simplifying the complex mixtures of peptides that are generated in typical proteomic analysis, where proteins are digested with trypsin and analyzed using liquid chromatography mass spectrometry data. The sequence-specific peptide selection not only reduces the complexity of digest mixtures, but also provides additional information for peptide identification. The targeted peptides are those that have either serine or threonine adjacent to a protease cleavage site. The sequence information should greatly aid in both database matching for protein identification and for de novo sequence determination.
Chelius
Dirk
D
Thermo Finnigan, 355 River Oaks Parkway, San Jose, California, 951 34, USA. dchelius@thrmofinnigan.com
Shaler
Thomas A
TA
D000345:Affinity Labels; D000447:Aldehydes; D000595:Amino Acid Sequence; D019921:Biotinylation; D002851:Chromatography, High Pressure Liquid; D013058:Mass Spectrometry; D010084:Oxidation-Reduction; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification*; D012694:Serine / Q000737:chemistry; D013912:Threonine / Q000737:chemistry
D000345:Affinity Labels; D000447:Aldehydes; D010455:Peptides; D013912:Threonine; D012694:Serine