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Catalytic properties of ADAM19
Valérie Chesneau1, J David Becherer, Yufang Zheng
1Cellular Biochemistry and Biophysics Program, Sloan-Kettering Institute, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
The Journal of Biological Chemistry
|April 19, 2003
Summary
ADAM19, a key protein in cardiovascular development, exhibits metalloprotease activity. It sheds tumor necrosis factor-related activation-induced cytokine (TRANCE) and regulates kit ligand-1 (KL-1) shedding.
Area of Science:
- Biochemistry
- Molecular Biology
- Developmental Biology
Background:
- ADAMs (a disintegrin and metalloproteinase domain) are glycoproteins involved in diverse biological processes, including development and ectodomain shedding.
- ADAM19 is crucial for cardiovascular morphogenesis, but its catalytic functions remain incompletely understood.
Purpose of the Study:
- To characterize the catalytic activity of soluble forms of ADAM19.
- To identify potential substrates and regulatory roles of ADAM19 in protein shedding.
Main Methods:
- Recombinant expression and purification of soluble ADAM19.
- In vitro proteolytic assays using myelin basic protein, insulin B chain, and specific peptide substrates.
- Enzyme inhibition assays using BB94 and TIMPs.
- Overexpression studies in COS-7 cells and analysis in mouse embryonic fibroblasts.
Main Results:
- Soluble ADAM19 demonstrated metalloprotease activity, evidenced by autocatalytic tag removal and cleavage of myelin basic protein and insulin B chain.
- ADAM19 activity was inhibited by BB94 but not by TIMPs 1-3.
- In vitro, ADAM19 cleaved substrates for TNF-alpha, TRANCE, and KL-1.
- ADAM19 overexpression increased TRANCE shedding, suggesting a role in osteoblasts.
- ADAM19 acted as a negative regulator of KL-1 shedding in tested cell lines.
Conclusions:
- Soluble ADAM19 possesses metalloprotease activity with specific substrate preferences.
- ADAM19 plays a role in TRANCE shedding and acts as a negative regulator of KL-1 shedding.
- These findings provide a basis for further investigation into ADAM19's cellular and in vivo functions.