Identification and Characterization of O-Biotinylated Hydroxy Amino Acid Residues in Peptides

✍ Scribed by B.T. Miller; M.E. Rogers; J.S. Smith; A. Kurosky

• Publisher: Elsevier Science
• Year: 1994
• Tongue: English
• Weight: 607 KB
• Volume: 219
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1006/abio.1994.1263

✦ Synopsis

Recent findings have indicated that certain hydroxyl groups of peptides have enhanced intrinsic chemical reactivity and can be O-acylated by N-hydroxysuccinimide esters of biotin. Analytical procedures are described for the identification of O-acylated derivatives of seryl, threonyl, and tyrosyl residues after reaction with the N-hydroxysuccinimide ester of biotin containing the extended spacer arm 6-aminohexanoic acid. The identification and chemical characterization of O-biotinylated residues included amino acid analysis, peptide sequence determination, and mass spectrometric analysis. The application of these analytical procedures provided unequivocal identification of O-biotinylated residues for a number of peptides investigated. In a separate study, we also show that O-biotinylated amino-terminal seryl residues occurring after proteolysis of peptides or proteins can rapidly undergo an O-->N acyl shift resulting in blockage of sequence analysis by the Edman degradation procedure.