Identification of Novel Natural Substrates of Fibroblast Activation Protein-alpha by Differential Degradomics and Proteomics

Identification of Novel Natural Substrates of Fibroblast Activation Protein-alpha by Differential Degradomics and Proteomics.

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Item Type: Article
Status: Published
Official URL: https://doi.org/10.1074/mcp.RA118.001046
Journal or Publication Title: Molecular & Cellular Proteomics
Volume: 18
Number: 1
Page Range: pp. 65-85
Date: 2018
Divisions: Liver Enzymes in Metabolism and Inflammation
Depositing User: General Admin
Identification Number: 10.1074/mcp.RA118.001046
ISSN: 1535-9476
Date Deposited: 21 Dec 2020 03:13
Abstract:

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.

Keywords: Coagulation; Cytokines; Degradomics; Endopeptidases; Extracellular Matrix; Fibroblast Activation Protein; Fibroblasts; Proteases; SILAC; Substrate Identification.

© 2019 Zhang et al.

Creators:
Creators
Email
Zhang, Hui Emma
UNSPECIFIED
Hamson, Elizabeth J.
UNSPECIFIED
Koczorowska, Maria Magdalena
UNSPECIFIED
Tholen, Stefan
UNSPECIFIED
Chowdhury, Sumaiya
UNSPECIFIED
Bailey, Charles G.
UNSPECIFIED
Lay, Angelina J.
UNSPECIFIED
Twigg, Stephen M.
UNSPECIFIED
Lee, Quintin
UNSPECIFIED
Roediger, Ben
UNSPECIFIED
Biniossek, Martin L.
UNSPECIFIED
O'Rourke, Matthew B.
UNSPECIFIED
McCaughan, Geoffrey W.
UNSPECIFIED
Keane, Fiona M.
UNSPECIFIED
Schilling, Oliver
UNSPECIFIED
Gorrell, Mark D.
UNSPECIFIED
Last Modified: 21 Dec 2020 03:13
URI: https://eprints.centenary.org.au/id/eprint/328

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