Post-translational regulation and proteolytic activity of the metalloproteinase ADAMTS8
| Autor: | Salvatore Santamaria, Josefin Ahnström, Kazuhiro Yamamoto, Daniel R. Martin, Xiangyi Dong, Suneel S. Apte |
|---|---|
| Přispěvatelé: | British Heart Foundation |
| Jazyk: | angličtina |
| Rok vydání: | 2021 |
| Předmět: |
PEI
polyethylenimine WT wild type osteopontin ADAMTS a disintegrin-like and metalloproteinase domain with thrombospondin motifs Biochemistry MEM minimum essential medium eagle ADAMTS Proteins Post-translational regulation 11 Medical and Health Sciences Aggrecanase versican Pulmonary Arterial Hypertension Pro prodomain biology Chemistry LC-MS/MS liquid chromatography–tandem mass spectrometry ADAMTS DMEM Dulbecco’s modified eagle’s medium Sp spacer LRP1 Cell biology ECM extracellular matrix ADAMTS4 TSR thrombospondin-like motif ADAMTS8 Proteoglycans PAH pulmonary arterial hypertension TIMP tissue inhibitor of metalloproteinase aggrecan 03 Chemical Sciences Research Article Proteases α2M alpha-2 macroglobulin Biochemistry & Molecular Biology alpha-2-Macroglobulin Humans TIMP Molecular Biology Dis disintegrin-like domain Tissue Inhibitor of Metalloproteinase-3 proteoglycan CR cysteine-rich domain LRP1 low-density lipoprotein receptor-related protein 1 Cell Biology 06 Biological Sciences MEF mouse embryonic fibroblast HEK293 Cells Mp metalloproteinase domain DTT dithiothreitol OPN osteopontin Proteolysis biology.protein PA-SMC pulmonary artery–smooth muscle cell Protein Processing Post-Translational |
| Zdroj: | JOURNAL OF BIOLOGICAL CHEMISTRY The Journal of Biological Chemistry |
| Popis: | A disintegrin-like and metalloprotease domain with thrombospondin type 1 motifs (ADAMTS)8 is a secreted protease, which was recently implicated in pathogenesis of pulmonary arterial hypertension (PAH). However, the substrate repertoire of ADAMTS8 and regulation of its activity are incompletely understood. Although considered a proteoglycanase because of high sequence similarity and close phylogenetic relationship to the proteoglycan-degrading proteases ADAMTS1, 4, 5, and 15, as well as tight genetic linkage with ADAMTS15 on human chromosome 11, its aggrecanase activity was reportedly weak. Several post-translational factors are known to regulate ADAMTS proteases such as autolysis, inhibition by endogenous inhibitors, and receptor-mediated endocytosis, but their impacts on ADAMTS8 are unknown. Here, we show that ADAMTS8 undergoes autolysis at six different sites within its spacer domain. We also found that in contrast to ADAMTS4 and 5, ADAMTS8 levels were not regulated through low-density lipoprotein receptor-related protein 1 (LRP1)-mediated endocytosis. Additionally, ADAMTS8 lacked significant activity against the proteoglycans aggrecan, versican, and biglycan. Instead, we found that ADAMTS8 cleaved osteopontin, a phosphoprotein whose expression is upregulated in PAH. Multiple ADAMTS8 cleavage sites were identified using liquid chromatography-tandem mass spectrometry. Osteopontin cleavage by ADAMTS8 was efficiently inhibited by TIMP-3, an endogenous inhibitor of ADAMTS1, 4, and 5, as well as by TIMP-2, which has no previously reported inhibitory activity against other ADAMTS proteases. These differences in post-translational regulation and substrate repertoire differentiate ADAMTS8 from other family members and may help to elucidate its role in PAH. |
| Databáze: | OpenAIRE |
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