期刊
MOLECULAR & CELLULAR PROTEOMICS
卷 18, 期 1, 页码 65-85出版社
AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC
DOI: 10.1074/mcp.RA118.001046
关键词
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资金
- Australian National Health and Medical Research Council [1105238]
- Rebecca L. Cooper Medical Research Foundation
- Australian Postgraduate Award
- Group of Eight Australia - Deutscher Akademischer Austauschdienst Joint Research Co-operation Scheme
- Deutsche Forschungsgemeinschaft [GR 1748/6-1, SCHI 871/8-1, SCHI 871/9-1, SCHI 871/11-1, SCHI 871/12-1, INST 39/900-1]
- Excellence Initiative of the German Federal and State Governments (EXC 294, BIOSS)
- Germany-Israel Foundation [I-1444-201.2/2017]
- Deutsche Forschungsgemeinschaft (SFB850-Project Z1) [INST 39/766-3]
Fibroblast activation protein-alpha (FAP) is a cell-surface transmembrane-anchored dimeric protease. This unique, constitutively active serine protease has both dipeptidyl aminopeptidase and endopeptidase activities and can hydrolyze the post-proline bond. FAP expression is very low in adult organs but is upregulated by activated fibroblasts in sites of tissue remodeling, including fibrosis, atherosclerosis, arthritis and tumors. To identify the endogenous substrates of FAP, we immortalized primary mouse embryonic fibroblasts (MEFs) from FAP gene knockout embryos and then stably transduced them to express either enzymatically active or inactive FAP. The MEF secretomes were then analyzed using degradomic and proteomic techniques. Terminal amine isotopic labeling of substrates (TAILS)-based degradomics identified cleavage sites in collagens, many other extracellular matrix (ECM) and associated proteins, and lysyl oxidase-like-1, CXCL-5, CSF-1, and C1qT6, that were confirmed in vitro. In addition, differential metabolic labeling coupled with quantitative proteomic analysis also implicated FAP in ECM-cell interactions, as well as with coagulation, metabolism and wound healing associated proteins. Plasma from FAP-deficient mice exhibited slower than wild-type clotting times. This study provides a significant expansion of the substrate repertoire of FAP and provides insight into the physiological and potential pathological roles of this enigmatic protease.
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