title: Autocatalytic processing of heme by lactoperoxidase produces the native protein-bound prosthetic group.
abstract: The covalently bound prosthetic group of lactoperoxidase (LPO) has been obtained by hydrolysis of the protein and identified as a dihydroxylated heme. A baculovirus expression system has been developed for LPO and used to obtain protein in which the heme is only partially covalently bound. Reaction of the purified heme. apoLPO complex with H2O2 results in both autocatalytic modification of the heme and covalent attachment to the protein. Hydrolytic experiments establish that the autocatalytically incorporated heme is bound normally. Two monohydroxylated heme intermediates have been detected. The peroxidative activity of LPO increases in proportion to the extent of covalently bound heme. The LPO results provide a paradigm for autocatalytic incorporation of heme groups into the mammalian peroxidases, including myeloperoxidase and eosinophil peroxidase, all of which exhibit strong sequence similarity with LPO and have covalently-bound heme groups.
authors.affiliation: Department of Pharmaceutical Chemistry, School of Pharmacy, University of California, San Francisco, California 94143-0446, USA.
authors.forename: G D
authors.initials: GD
authors.lastname: DePillis
authors.forename: S i
authors.initials: Si
authors.lastname: Ozaki
authors.forename: J M
authors.initials: JM
authors.lastname: Kuo
authors.forename: D A
authors.initials: DA
authors.lastname: Maltby
authors.forename: P R
authors.initials: PR
authors.lastname: Ortiz de Montellano
mesh_terms: D000818:Animals; D002384:Catalysis; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D018076:DNA, Complementary / Q000737:chemistry; D006418:Heme / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D006868:Hydrolysis; D007784:Lactoperoxidase / Q000378:metabolism*; D008956:Models, Chemical; D009195:Peroxidase / Q000378:metabolism; D011402:Pronase / Q000378:metabolism; D011994:Recombinant Proteins / Q000032:analysis
chemical_list: D018076:DNA, Complementary; D011994:Recombinant Proteins; D006418:Heme; D006861:Hydrogen Peroxide; D007784:Lactoperoxidase; D009195:Peroxidase; D011402:Pronase