Imperial College London
Alternate

ProfessorJimCrawley

Faculty of Medicine, Department of Immunology and Inflammation

Professor of Haemostasis
 
 
 
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Contact

 

+44 (0)20 3313 2297j.crawley Website

 
 
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Location

 

5S5aCommonwealth BuildingHammersmith Campus

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Summary

 

Publications

Citation

BibTex format

@article{Colige:2019:10.1074/jbc.RA119.007492,
author = {Colige, A and Monseur, C and Crawley, J and Santamaria, S and de, Groot R},
doi = {10.1074/jbc.RA119.007492},
journal = {Journal of Biological Chemistry},
pages = {8037--8045},
title = {Proteomic discovery of substrates of the cardiovascular protease ADAMTS7},
url = {http://dx.doi.org/10.1074/jbc.RA119.007492},
volume = {294},
year = {2019}
}
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RIS format (EndNote, RefMan)

TY  - JOUR
AB  - The protease ADAMTS7 functions in the extracellular matrix (ECM) of the cardiovascular system. However, its physiological substrate specificity and mechanism of regulation remain to be explored. To address this, we conducted an unbiased substrate analysis using terminal amine isotopic labeling of substrates (TAILS). The analysis identified candidate substrates of ADAMTS7 in the human fibroblast secretome, including proteins with a wide range of functions, such as collagenous and noncollagenous extracellular matrix proteins, growth factors, proteases, and cell-surface receptors. It also suggested that autolysis occurs at Glu-729–Val-730 and Glu-732–Ala-733 in the ADAMTS7 Spacer domain, which was corroborated by N-terminal sequencing and Western blotting. Importantly, TAILS also identified proteolysis of the latent TGF-β–binding proteins 3 and 4 (LTBP3/4) at a Glu-Val and Glu-Ala site, respectively. Using purified enzyme and substrate, we confirmed ADAMTS7-catalyzed proteolysis of recombinant LTBP4. Moreover, we identified multiple additional scissile bonds in an N-terminal linker region of LTBP4 that connects fibulin-5/tropoelastin and fibrillin-1–binding regions, which have an important role in elastogenesis. ADAMTS7-mediated cleavage of LTBP4 was efficiently inhibited by the metalloprotease inhibitor TIMP-4, but not by TIMP-1 and less efficiently by TIMP-2 and TIMP-3. As TIMP-4 expression is prevalent in cardiovascular tissues, we propose that TIMP-4 represents the primary endogenous ADAMTS7 inhibitor. In summary, our findings reveal LTBP4 as an ADAMTS7 substrate, whose cleavage may potentially impact elastogenesis in the cardiovascular system. We also identify TIMP-4 as a likely physiological ADAMTS7 inhibitor.
AU  - Colige,A
AU  - Monseur,C
AU  - Crawley,J
AU  - Santamaria,S
AU  - de,Groot R
DO  - 10.1074/jbc.RA119.007492
EP  - 8045
PY  - 2019///
SN  - 0021-9258
SP  - 8037
TI  - Proteomic discovery of substrates of the cardiovascular protease ADAMTS7
T2  - Journal of Biological Chemistry
UR  - http://dx.doi.org/10.1074/jbc.RA119.007492
UR  - http://hdl.handle.net/10044/1/69856
VL  - 294
ER  -
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