Expression of endothelin-converting enzyme in both neuroblastoma and glial cell lines and its localization in rat hippocampus.

Abstract

Endothelin-converting enzyme is a phosphoramidon-sensitive metalloprotease that cleaves big endothelin to the potent vasoconstrictor peptide, endothelin. The converting enzyme is expressed in endothelial cells in a variety of tissues and in some secretory cells. In the present study, phosphoramidon-sensitive endothelin-converting enzyme activity has been demonstrated by radioimmunoassay in the neuroblastoma cell line, SH-SY5Y, and in Bu17 and C6 glioma lines. The identity of the activity was confirmed by immunoblotting, revealing a polypeptide of approximately 120 kDa in each of these lines, in D384 glioma cells, and in primary astrocytes. Immunofluorescence revealed the cell-surface location of endothelin-converting enzyme in the neuronal and glial cell lines and in primary astrocytes. Pretreatment of SH-SY5Y and Bu17 cells with phosphoramidon resulted in an apparent concentration of the enzyme protein in an intracellular compartment. Immunoperoxidase-staining of rat brain sections located this metalloprotease to the pyramidal cells of the hippocampus. Endothelin-converting enzyme-1 was revealed by in situ hybridisation in the neuronal and glial cell lines.

• Authors: K Barnes, B J Walkden, T C Wilkinson, A J Turner
• Journal: Journal of neurochemistry (J Neurochem) Vol. 68 Issue 2 Pg. 570-7 (Feb 1997) ISSN: 0022-3042 [Print] England
• PMID: 9003042 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antibodies, Monoclonal
• DNA, Complementary
• Glial Fibrillary Acidic Protein
• Glycopeptides
• Protease Inhibitors
• RNA, Messenger
• CD13 Antigens
• Aspartic Acid Endopeptidases
• Metalloendopeptidases
• Neprilysin
• Endothelin-Converting Enzymes
• phosphoramidon

Topics

• Animals
• Antibodies, Monoclonal
• Aspartic Acid Endopeptidases (genetics, immunology, metabolism)
• Astrocytes (chemistry, enzymology)
• CD13 Antigens (analysis, metabolism)
• Cell Membrane (chemistry)
• Cells, Cultured (chemistry, enzymology)
• Cloning, Molecular
• DNA, Complementary
• Endothelin-Converting Enzymes
• Fluorescent Antibody Technique
• Gene Expression Regulation, Enzymologic (drug effects, physiology)
• Glial Fibrillary Acidic Protein (analysis)
• Glioma
• Glycopeptides (pharmacology)
• Golgi Apparatus (chemistry)
• Hippocampus (enzymology)
• Immunoenzyme Techniques
• In Situ Hybridization
• Metalloendopeptidases (genetics, immunology, metabolism)
• Neprilysin (analysis, metabolism)
• Neuroblastoma
• Neuroglia (enzymology)
• Protease Inhibitors (pharmacology)
• RNA, Messenger (analysis)
• Rats
• Tumor Cells, Cultured (enzymology)