title: Ion-selective enrichment of tyrosine-sulfated peptides from complex protein digests.
abstract: We have developed a novel procedure for concentrating sulfated peptides, as a front end to mass spectrometric analysis, based on ion-selective interaction of sulfate ions with anion exchangers. Ions with a higher charge and smaller solvated ion radius, such as sulfate ions, have higher retention in an ion exchanger due to their greater degree of coulombic interactions. We tested the effectiveness of this approach for enrichment and identification of sulfated peptides using a tryptic digest of bovine serum albumin spiked with model sulfated peptide (molar ratio 20:1) and using a tryptic digest of bovine fibrinogen. Sulfated peptides are identified by mass spectrometry in which both the molecular ion and its specific fragment ion produced by facile loss of SO(3) are detected. In both experiments, sulfated peptides were strongly retained on the anion exchanger and were eluted by higher concentrations of competing ion with minimal contamination of nonsulfated peptides. Using this procedure, we determined that the 13-amino acid C-terminal peptide of the minor gamma'-chain of bovine fibrinogen contains sulfated tyrosine.
authors.affiliation: Graduate School of Bio-Agricultural Sciences, Nagoya University, Chikusa, Japan.
authors.forename: Yukari
authors.initials: Y
authors.lastname: Amano
authors.forename: Hidefumi
authors.initials: H
authors.lastname: Shinohara
authors.forename: Youji
authors.initials: Y
authors.lastname: Sakagami
authors.forename: Yoshikatsu
authors.initials: Y
authors.lastname: Matsubayashi
mesh_terms: D000595:Amino Acid Sequence; D002852:Chromatography, Ion Exchange; D005340:Fibrinogen / Q000032:analysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis*; D011499:Protein Processing, Post-Translational; D013431:Sulfates / Q000378:metabolism; D014443:Tyrosine / Q000378:metabolism*
chemical_list: D010455:Peptides; D013431:Sulfates; D014443:Tyrosine; D005340:Fibrinogen