title: Multidomain human peroxidasin 1 is a highly glycosylated and stable homotrimeric high spin ferric peroxidase.
abstract: Human peroxidasin 1 (hsPxd01) is a multidomain heme peroxidase that uses bromide as a cofactor for the formation of sulfilimine cross-links. The latter confers critical structural reinforcement to collagen IV scaffolds. Here, hsPxd01 and various truncated variants lacking nonenzymatic domains were recombinantly expressed in HEK cell lines. The N-glycosylation site occupancy and disulfide pattern, the oligomeric structure, and unfolding pathway are reported. The homotrimeric iron protein contains a covalently bound ferric high spin heme per subunit with a standard reduction potential of the Fe(III)/Fe(II) couple of -233 ± 5 mV at pH 7.0. Despite sequence homology at the active site and biophysical properties similar to human peroxidases, the catalytic efficiency of bromide oxidation (kcat/KM(app)) of full-length hsPxd01 is rather low but increased upon truncation. This is discussed with respect to its structure and proposed biosynthetic function in collagen IV cross-linking.
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Monika
authors.initials: M
authors.lastname: Soudi
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Martina
authors.initials: M
authors.lastname: Paumann-Page
authors.affiliation: the Laboratory of Pharmaceutical Chemistry and Analytical Platform of the Faculty of Pharmacy, Faculty of Pharmacy, Université Libre de Bruxelles, 1050 Brussels, Belgium.
authors.forename: Cedric
authors.initials: C
authors.lastname: Delporte
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Katharina F
authors.initials: KF
authors.lastname: Pirker
authors.affiliation: the Departments of Life Sciences and.
authors.forename: Marzia
authors.initials: M
authors.lastname: Bellei
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Eva
authors.initials: E
authors.lastname: Edenhofer
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Stadlmayr
authors.affiliation: Chemistry and Geology, University of Modena and Reggio Emilia, 41125 Modena, Italy, and.
authors.forename: Gianantonio
authors.initials: G
authors.lastname: Battistuzzi
authors.affiliation: the Laboratory of Experimental Medicine (ULB 222 Unit), CHU de Charleroi, A. Vésale Hospital, Université Libre de Bruxelles, 6110 Montigny-le-Tilleul, Belgium.
authors.forename: Karim Zouaoui
authors.initials: KZ
authors.lastname: Boudjeltia
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria.
authors.forename: Paul G
authors.initials: PG
authors.lastname: Furtmüller
authors.affiliation: the Laboratory of Pharmaceutical Chemistry and Analytical Platform of the Faculty of Pharmacy, Faculty of Pharmacy, Université Libre de Bruxelles, 1050 Brussels, Belgium.
authors.forename: Pierre
authors.initials: P
authors.lastname: Van Antwerpen
authors.affiliation: From the Department of Chemistry, Division of Biochemistry, BOKU-University of Natural Resources and Life Sciences, Muthgasse 18, A-1190 Vienna, Austria, christian.obinger@boku.ac.at.
authors.forename: Christian
authors.initials: C
authors.lastname: Obinger
mesh_terms: D000951:Antigens, Neoplasm / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D002384:Catalysis; D024141:Collagen Type IV / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D057809:HEK293 Cells; D006801:Humans; D007501:Iron / Q000737:chemistry* / Q000378:metabolism; D010084:Oxidation-Reduction; D010544:Peroxidases; D017434:Protein Structure, Tertiary; D017472:Receptors, Interleukin-1 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D013329:Structure-Activity Relationship; D000097423:Peroxidasin
chemical_list: D000951:Antigens, Neoplasm; D024141:Collagen Type IV; D007501:Iron; D010544:Peroxidases; D017472:Receptors, Interleukin-1; C535215:PXDN protein, human; D000097423:Peroxidasin
keywords: Collagen; Extracellular Matrix Protein; Glycosylation; Metalloenzyme; Peroxidase; Peroxidasin; Vascular Peroxidase