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

Benjamin Sevcnikar, Irene Schaffner, Christine Y. Chuang, Luke Gamon, Martina Paumann-Page, Stefan Hofbauer, Michael J. Davies, Paul G. Furtmüller, Christian Obinger*

*Corresponding author for this work

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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 (S[dbnd]N) 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.

Original languageEnglish
Article number108443
JournalArchives of Biochemistry and Biophysics
Volume689
Number of pages7
ISSN0003-9861
DOIs
Publication statusPublished - 15 Aug 2020

Keywords

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

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