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Characterization of METH-1/ADAMTS1 processing reveals two distinct active forms.
J C Rodriguez-Manzaneque1, A B Milchanowski, E K Dufour
1Department of Molecular, Cell, and Developmental Biology and Molecular Biology Institute, UCLA, Los Angeles, California 90095, USA.
The Journal of Biological Chemistry
|August 17, 2000
Summary
The METH-1/ADAMTS1 protein undergoes sequential processing, generating active forms that modulate its anti-angiogenic properties. This processing impacts endothelial cell proliferation and protein interactions.
Area of Science:
- Biochemistry
- Molecular Biology
- Cell Biology
Background:
- METH-1/ADAMTS1 is a metalloprotease with anti-angiogenic properties.
- The protein contains metalloprotease, disintegrin, and thrombospondin-like motifs.
Purpose of the Study:
- To investigate the processing of secreted human pro-METH-1.
- To determine the functional consequences of METH-1 processing on its anti-angiogenic activity.
Main Methods:
- In vitro processing assays using purified furin and matrix metalloproteases (MMPs).
- Inhibition studies with specific protease inhibitors.
- Analysis of protein processing products and their interaction with heparin and endothelial cells.
Main Results:
- Pro-METH-1 is processed in two steps to yield p87 and p65 active forms.
- Furin generates p87, while MMPs (MMP-2, -8, -15) generate p65 from p87.
- The p65 form, lacking C-terminal thrombospondin repeats, shows reduced affinity for heparin and endothelial cells, and decreased anti-proliferative activity.
Conclusions:
- METH-1/ADAMTS1 processing by furin and MMPs generates distinct active fragments.
- This processing regulates METH-1's anti-angiogenic activity by altering its functional properties.
- The sequential processing is crucial for modulating METH-1's biological effects.