TY - JOUR

T1 - The leucine-rich repeat domain of human peroxidasin 1 promotes binding to laminin in basement membranes

AU - Sevcnikar, Benjamin

AU - Schaffner, Irene

AU - Chuang, Christine Y.

AU - Gamon, Luke

AU - Paumann-Page, Martina

AU - Hofbauer, Stefan

AU - Davies, Michael J.

AU - Furtmüller, Paul G.

AU - Obinger, Christian

PY - 2020/8/15

Y1 - 2020/8/15

N2 - 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.

AB - 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.

KW - Basement membrane

KW - Extracellular matrix

KW - Human peroxidasin 1

KW - Immunoprecipitation

KW - Laminin

KW - Surface plasmon resonance spectroscopy

KW - Type-IV collagen

UR - http://www.scopus.com/inward/record.url?scp=85086475596&partnerID=8YFLogxK

U2 - 10.1016/j.abb.2020.108443

DO - 10.1016/j.abb.2020.108443

M3 - Journal article

C2 - 32485152

AN - SCOPUS:85086475596

VL - 689

JO - Archives of Biochemistry and Biophysics

JF - Archives of Biochemistry and Biophysics

SN - 0003-9861

M1 - 108443

ER -