title: Characterization of tyrosine sulfate residues in antihemophilic recombinant factor VIII by liquid chromatography electrospray ionization tandem mass spectrometry and amino acid analysis.
abstract: Recombinant Factor VIII (rFVIII) is involved in the cascade of biochemical reactions leading to blood coagulation and is used for the treatment of haemophilia A. Plasma-derived FVIII (pdFVIII) has been reported to be post-translationally modified by sulfation of tyrosine residues at positions 346, 1664, 1680, 718, 719 and 723. This report describes the quantitation of tyrosine sulfate residues in BHK-derived, human rFVIII by amino acid composition analysis and the identification of their positions in the polypeptide sequence using a combination of liquid chromatography and electrospray ionization mass spectrometry in the analysis of proteolytic digests of the protein.
authors.affiliation: Bayer Pharmaceuticals, Berkeley, CA 94701, USA.
authors.forename: J C
authors.initials: JC
authors.lastname: Severs
authors.forename: M
authors.initials: M
authors.lastname: Carnine
authors.forename: H
authors.initials: H
authors.lastname: Eguizabal
authors.forename: K K
authors.initials: KK
authors.lastname: Mock
mesh_terms: D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D002851:Chromatography, High Pressure Liquid; D005169:Factor VIII / Q000032:analysis*; D006868:Hydrolysis; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000032:analysis; D010449:Peptide Mapping; D011994:Recombinant Proteins / Q000032:analysis; D013056:Spectrophotometry, Ultraviolet; D013917:Thrombin / Q000737:chemistry; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000032:analysis
chemical_list: D000596:Amino Acids; D010446:Peptide Fragments; D011994:Recombinant Proteins; C005662:tyrosine O-sulfate; D014443:Tyrosine; D005169:Factor VIII; D013917:Thrombin