Peroxidasin-mediated crosslinking of collagen IV is independent of NADPH oxidases

Sirokmány, Gábor; Kovács, Hajnal A.; Lázár, Enikő; Kónya, Krisztina; Donkó, Ágnes; Enyedi, Balázs; Grasberger, Helmut; Geiszt, Miklós

doi:10.1016/j.redox.2018.03.009

Cited by 18 publications

(17 citation statements)

References 40 publications

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“…11,34 Crucial to the stable formation of these bonds are sulfilamine bonds, and these are catalyzed in a peroxidase reaction by peroxidasin, or PXDN. 35 This may explain why the phenotype associated with PXDN may be so severe, as demonstrated in case 21 (Fig. 2c) of this study, and as shown in the mouse pxdn nonsense variant model.…”

Section: Variants In Collagen and Extracellular Matrix-associated Gensupporting

confidence: 60%

Revealing hidden genetic diagnoses in the ocular anterior segment disorders

Yousoof

Grigg

et al. 2020

Genetics in Medicine

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Purpose: Ocular anterior segment disorders (ASDs) are clinically and genetically heterogeneous, and genetic diagnosis often remains elusive. In this study, we demonstrate the value of a combined analysis protocol using phenotypic, genomic, and pedigree structure data to achieve a genetic conclusion. Methods: We utilized a combination of chromosome microarray, exome sequencing, and genome sequencing with structural variant and trio analysis to investigate a cohort of 41 predominantly sporadic cases. Results: We identified likely causative variants in 54% (22/41) of cases, including 51% (19/37) of sporadic cases and 75% (3/4) of cases initially referred as familial ASD. Two-thirds of sporadic cases were found to have heterozygous variants, which in most cases were de novo. Approximately one-third (7/22) of genetic diagnoses were found in rarely reported or recently identified ASD genes including PXDN, GJA8, COL4A1, ITPR1, CPAMD8, as well as the new phenotypic association of Axenfeld-Rieger anomaly with a homozygous ADAMTS17 variant. The remainder of the variants were in key ASD genes including FOXC1, PITX2, CYP1B1, FOXE3, and PAX6. Conclusions: We demonstrate the benefit of detailed phenotypic, genomic, variant, and segregation analysis to uncover some of the previously "hidden" heritable answers in several rarely reported and newly identified ocular ASD-related disease genes.

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Section: Variants In Collagen and Extracellular Matrix-associated Gensupporting

confidence: 60%

Revealing hidden genetic diagnoses in the ocular anterior segment disorders

Yousoof

Grigg

et al. 2020

Genetics in Medicine

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“…Such PXDN functions require both the ECM motifs and peroxidase activity, which can be blocked by anti-PXDN antibody and peroxidase inhibitor by NOX enzymes. 4,27 The low enzymatic activity of PXDN led to the hypothesis that the PXDN in myofibroblasts is involved in a novel ECM-formation pathway that is not mediated by peroxidase activity. 7 Our study showed, however, that PXDN functions extracellularly as a secreted protein and that its functional loss in ECs is driven by deletions of the peroxidase domain or by peroxidase inhibitor.…”

Section: Discussionmentioning

confidence: 99%

Peroxidasin is essential for endothelial cell survival and growth signaling by sulfilimine crosslink‐dependent matrix assembly

et al. 2020

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Peroxidasin (PXDN) has been reported to crosslink the C-terminal non-collagenous domains of collagen IV (Col IV) by forming covalent sulfilimine bond. Here, we explored the physiological role of PXDN and its mechanism of action in endothelial cell survival and growth. Silencing of PXDN using siRNAs decreased cell proliferation without increase of the number of detached cells and decreased cell viability under serum-starved condition with increased fragmented nuclei and caspase 3/7 activity. Conditioned medium (CM) containing wild-type PXDN restored the proliferation of PXDN-depleted cells, but CM containing mutant PXDN with deletion of either N-terminal extracellular matrix (ECM) motifs or peroxidase domain failed to restore PXDN function. Accordingly, anti-PXDN antibody [raised against IgC2 (3-4) subdomain within ECM motifs] and peroxidase inhibitor phloroglucinol prevented the rescue of the PXDN-depleted cells by PXDN-containing CM. PXDN depletion resulted in loss of sulfilimine crosslinks, and decreased dense fibrillar network assembly of not only Col IV, but also fibronectin and laminin like in Col IV knockdown. Exogenous PXDN-containing CM restored ECM assembly as well as proliferation of PXDN-depleted cells. Accordingly, purified recombinant PXDN protein restored the proliferation and ECM assembly, and prevented cell death of the PXDN-depleted cells. PXDN depletion also showed reduced growth factors-induced phosphorylation of FAK and ERK1/2. In addition, siPXDN-transfected cell-derived matrix failed to provide full ECM-mediated activation of FAK and ERK1/2. These results indicate that both the ECM motifs and peroxidase activity are essential for the cellular | 10229

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“…We assessed the level of NC1 domain sulfilimine crosslinking in Col IV according to a procedure reported previously [32]. Briefly, the lung tissues were lysed in hypotonic lysis buffer (10 mM CaCl 2 , 50 mM HEPES, pH 7.4, 0.1 mM benzamininde hydrochloride, 25 mM 6-aminocaproic acid, and 1 mM PMSF).…”

Section: Detection Of Nc1 Sulfilimine Crosslinkmentioning

confidence: 99%

Biallelic Deletion of Pxdn in Mice Leads to Anophthalmia and Severe Eye Malformation

Kim

Ham

Lee

et al. 2019

IJMS

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Peroxidasin (PXDN) is a unique peroxidase containing extracellular matrix motifs and stabilizes collagen IV networks by forming sulfilimine crosslinks. PXDN gene knockout in Caenorhabditis elegans (C. elegans) and Drosophila results in the demise at the embryonic and larval stages. PXDN mutations lead to severe eye disorders, including microphthalmia, cataract, glaucoma, and anterior segment dysgenesis in humans and mice. To investigate how PXDN loss of function affects organ development, we generated Pxdn knockout mice by deletion of exon 1 and its 5′ upstream sequences of the Pxdn gene using the CRISPR/Cas9 system. Loss of both PXDN expression and collagen IV sulfilimine cross-links was detected only in the homozygous mice, which showed completely or almost closed eyelids with small eyes, having no apparent external morphological defects in other organs. In histological analysis of eye tissues, the homozygous mice had extreme defects in eye development, including no eyeballs or drastically disorganized eye structures, whereas the heterozygous mice showed normal eye structure. Visual function tests also revealed no obvious functional abnormalities in the eyes between heterozygous mice and wild-type mice. Thus, these results suggest that PXDN activity is essential in eye development, and also indicate that a single allele of Pxdn gene is sufficient for eye-structure formation and normal visual function.

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Peroxidasin-mediated crosslinking of collagen IV is independent of NADPH oxidases

Cited by 18 publications

References 40 publications

Revealing hidden genetic diagnoses in the ocular anterior segment disorders

Revealing hidden genetic diagnoses in the ocular anterior segment disorders

Peroxidasin is essential for endothelial cell survival and growth signaling by sulfilimine crosslink‐dependent matrix assembly

Biallelic Deletion of Pxdn in Mice Leads to Anophthalmia and Severe Eye Malformation

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