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Endothelin-converting enzyme: ultrastructural localization and its recycling from the cell surface
K Barnes1, C Brown, A J Turner
1Department of Biochemistry and Molecular Biology, University of Leeds, UK. k.barnes@leeds.ac.uk
Hypertension (Dallas, Tex. : 1979)
|February 4, 1998
Summary
Endothelial cells generate endothelin-1 (ET-1) via endothelin-converting enzyme (ECE). This study reveals ECE
Area of Science:
- Endothelial cell biology
- Cardiovascular research
- Enzymology
Background:
- Endothelin-1 (ET-1) is a potent vasoconstrictor involved in cardiovascular diseases.
- ET-1 is produced from big ET-1 by endothelin-converting enzyme (ECE).
- The cellular localization and trafficking of ECE are not fully understood.
Purpose of the Study:
- To determine the subcellular localization of ECE in endothelial cells.
- To investigate the cell surface expression and recycling of ECE.
- To colocalize ECE with big ET-1 and other relevant proteins.
Main Methods:
- Ultrastructural localization using immunogold labeling.
- Double immunogold staining on ultrathin cryosections.
- Cell surface biotinylation assays.
- Immunofluorescence staining after chloroquine treatment.
Main Results:
- ECE is localized to the cell surface and intracellular vesicles in endothelial cells.
- ECE exhibits cell surface recycling.
- First ultrastructural localization of ECE in lung tissue.
- Colocalization of big ET-1 and ECE in 50-100 nm vesicles.
- ECE and angiotensin-converting enzyme found on the luminal membrane.
- ECE confirmed on the plasma membrane using biotinylation.
- ECE and trans-Golgi network 38 protein share an intracellular compartment.
Conclusions:
- ECE is dynamically localized on the cell surface and intracellular compartments of endothelial cells.
- ECE trafficking and localization are crucial for ET-1 regulation.
- This study provides novel insights into ECE's role in endothelial function and cardiovascular pathophysiology.