title: The status of half-cystine residues and locations of N-glycosylated asparagine residues in human eosinophil peroxidase.
abstract: The determination by protein chemistry methods of the half-cystine status in human eosinophil peroxidase (EPO) is reported. EPO is two-chained and has a total of 14 half-cystine residues. Cys141 and Cys152 form an intrachain bridge in the light chain of EPO. Disulfide bridges connect Cys253 and Cys263, Cys257 and Cys287, Cys359 and Cys370, Cys570 and Cys635, and Cys676 and Cys701, forming five intrachain disulfide bridges in the heavy chain of EPO. Cys291 and Cys455 are found to be unpaired, containing free sulfhydryl groups. The pattern of disulfide bridges is in agreement with that predicted from the X-ray crystallographic structure of canine myeloperoxidase (MPO) (Zeng, J., and Fenna, R. E. (1992) J. Mol. Biol. 226, 185-207) to be general for the class of mammalian peroxidases, including EPO, MPO, lactoperoxidase (LPO), and thyroid peroxidase (TPO). Of four candidate sites in EPO for attachment of glucosamine-based carbohydrate, Asn327 and Asn363 are occupied, whereas Asn700 and Asn708 are unsubstituted. Furthermore, a discrepancy in the literature regarding the sequence of residues 645-659 is resolved.
authors.affiliation: Department of Molecular and Structural Biology, University of Aarhus, Science Park, Gustav Wieds Vej 10 C, Arhus C, DK-8000, Denmark.
authors.forename: A R
authors.initials: AR
authors.lastname: Thomsen
authors.forename: L
authors.initials: L
authors.lastname: Sottrup-Jensen
authors.forename: G J
authors.initials: GJ
authors.lastname: Gleich
authors.forename: C
authors.initials: C
authors.lastname: Oxvig
mesh_terms: D000595:Amino Acid Sequence; D001216:Asparagine / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid; D003553:Cystine / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D047209:Eosinophil Peroxidase; D004804:Eosinophils / Q000201:enzymology*; D006023:Glycoproteins / Q000737:chemistry; D006031:Glycosylation; D006801:Humans; D008969:Molecular Sequence Data; D010544:Peroxidases / Q000737:chemistry*; D020539:Sequence Analysis, Protein
chemical_list: D004220:Disulfides; D006023:Glycoproteins; D003553:Cystine; D001216:Asparagine; D047209:Eosinophil Peroxidase; D010544:Peroxidases