Peroxidasin is essential for endothelial cell survival and growth signaling by sulfilimine crosslink-dependent matrix assembly
- 주제(기타) Biochemistry & Molecular Biology
- 주제(기타) Biology
- 주제(기타) Cell Biology
- 설명문(일반) [Lee, Seung-Woo; Kim, Hyun-Kyung; Naidansuren, Purevjargal; Ham, Kyung A.; Choi, Hong Seok; Joe, Young Ae] Catholic Univ Korea, Canc Res Inst, Coll Med, 222 Banpo Daero, Seoul 06519, South Korea; [Lee, Seung-Woo; Kim, Hyun-Kyung; Naidansuren, Purevjargal; Ham, Kyung A.; Choi, Hong Seok; Joe, Young Ae] Catholic Univ Korea, Coll Med, Dept Med Life Sci, 222 Banpo Daero, Seoul 06519, South Korea; [Lee, Seung-Woo; Kim, Hyun-Kyung; Naidansuren, Purevjargal; Ham, Kyung A.; Choi, Hong Seok; Joe, Young Ae] Catholic Univ Korea, Coll Med, Canc Evolut Res Ctr, Seoul, South Korea; [Ham, Kyung A.; Choi, Hong Seok; Joe, Young Ae] Catholic Univ Korea, Dept Biomed & Hlth Sci, Seoul, South Korea; [Ahn, Hyun-Young] Catholic Univ Korea, Coll Med, Dept Gynecol, Div Maternal Fetal Med, Seoul, South Korea; [Kim, Myungshin] Catholic Univ Korea, Coll Med, Dept Lab Med, Seoul, South Korea; [Kang, Dong Hoon; Kang, Sang Won] Ewha Womans Univ, Dept Life Sci, Seoul, South Korea; [Kang, Dong Hoon; Kang, Sang Won] Ewha Womans Univ, Res Ctr Cell Homeostasis, Seoul, South Korea
- 등재 SCIE, SCOPUS
- OA유형 Bronze
- 발행기관 WILEY
- 발행년도 2020
- 총서유형 Journal
- URI http://www.dcollection.net/handler/ewha/000000174978
- 본문언어 영어
- Published As http://dx.doi.org/10.1096/fj.201902899R
- PubMed https://pubmed.ncbi.nlm.nih.gov/32543734
초록/요약
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 function of PXDN and that PXDN is crucial for ECM assembly for survival and growth signaling.
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