2. Academic Validation
  3. The leucine-rich repeat domain of human peroxidasin 1 promotes binding to laminin in basement membranes

The leucine-rich repeat domain of human peroxidasin 1 promotes binding to laminin in basement membranes

  • Arch Biochem Biophys. 2020 Aug 15;689:108443. doi: 10.1016/j.abb.2020.108443.
Benjamin Sevcnikar 1,  Irene Schaffner 1,  Christine Y Chuang 2,  Luke Gamon 2,  Martina Paumann-Page 3,  Stefan Hofbauer 1,  Michael J Davies 2,  Paul G Furtmüller 4,  Christian Obinger 5
Affiliations

Affiliations

  • 1 Department of Chemistry, Institute of Biochemistry, BOKU - University of Natural Resources and Life Sciences, Muthgasse 18, A-1190, Vienna, Austria.
  • 2 Department of Biomedical Sciences, University of Copenhagen, Copenhagen, Denmark.
  • 3 Department of Pathology and Biomedical Science, University of Otago, Christchurch, New Zealand.
  • 4 Department of Chemistry, Institute of Biochemistry, BOKU - University of Natural Resources and Life Sciences, Muthgasse 18, A-1190, Vienna, Austria. Electronic address: paul.furtmueller@boku.ac.at.
  • 5 Department of Chemistry, Institute of Biochemistry, BOKU - University of Natural Resources and Life Sciences, Muthgasse 18, A-1190, Vienna, Austria. Electronic address: christian.obinger@boku.ac.at.
PMID: 32485152 DOI: 10.1016/j.abb.2020.108443
Abstract

Human peroxidasin 1 (PXDN) is a homotrimeric multidomain heme peroxidase and essential for tissue development and architecture. It has a biosynthetic function and catalyses the hypobromous acid-mediated formation of specific covalent sulfilimine (SN) bonds, which cross-link type IV collagen chains in basement membranes. Currently, it is unknown whether and which domain(s) [i.e. leucine-rich repeat domain (LRR), immunoglobulin domains, peroxidase domain, von Willebrand factor type C domain] of PXDN interact with the polymeric networks of the extracellular matrix (ECM), and how these interactions integrate and regulate the enzyme's cross-linking activity, without imparting oxidative damage to the ECM. In this study, we probed the interactions of four PXDN constructs with different domain compositions with components of a basement membrane extract by immunoprecipitation. Strong binding of the LRR-containing construct was detected with the major ECM protein laminin. Analysis of these interactions by surface plasmon resonance spectroscopy revealed similar kinetics and affinities of binding of the LRR-containing construct to human and murine laminin-111, with calculated dissociation constants of 1.0 and 1.5 μM, respectively. The findings are discussed with respect to the recently published in-solution structures of the PXDN constructs and the proposed biological role of this peroxidase.

Keywords

Basement membrane; Extracellular matrix; Human peroxidasin 1; Immunoprecipitation; Laminin; Surface plasmon resonance spectroscopy; Type-IV collagen.

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