Identification and characterization of O-biotinylated hydroxy amino acid residues in peptides.
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.
Miller
B T
BT
Department of Anatomy and Neurosciences, University of Texas Medical Branch, Galveston 77555.
Rogers
M E
ME
Smith
J S
JS
Kurosky
A
A
D000595:Amino Acid Sequence; D000682:Amyloid / Q000737:chemistry*; D001710:Biotin; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D002918:Chymotrypsin; D006801:Humans; D019811:Hydroxylamine; D006898:Hydroxylamines; D007202:Indicators and Reagents; D058228:Islet Amyloid Polypeptide; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry* / Q000302:isolation & purification; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides / Q000737:chemistry* / Q000302:isolation & purification; D012694:Serine; D016339:Spectrometry, Mass, Fast Atom Bombardment / Q000379:methods; D013912:Threonine
D000682:Amyloid; D006898:Hydroxylamines; D007202:Indicators and Reagents; D058228:Islet Amyloid Polypeptide; D009842:Oligopeptides; D010446:Peptide Fragments; D010455:Peptides; D019811:Hydroxylamine; D013912:Threonine; D012694:Serine; D001710:Biotin; D002918:Chymotrypsin