Expression of endothelin-converting enzyme in both neuroblastoma and glial cell lines and its localization in rat hippocampus.
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.
Barnes
K
K
Department of Biochemistry and Molecular Biology, University of Leeds, England.
Walkden
B J
BJ
Wilkinson
T C
TC
Turner
A J
AJ
D000818:Animals; D000911:Antibodies, Monoclonal; D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D001253:Astrocytes / Q000737:chemistry / Q000201:enzymology; D018826:CD13 Antigens / Q000032:analysis / Q000378:metabolism; D002462:Cell Membrane / Q000737:chemistry; D002478:Cells, Cultured / Q000737:chemistry / Q000201:enzymology; D003001:Cloning, Molecular; D018076:DNA, Complementary; D000072396:Endothelin-Converting Enzymes; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects / Q000502:physiology; D005904:Glial Fibrillary Acidic Protein / Q000032:analysis; D005910:Glioma; D006020:Glycopeptides / Q000494:pharmacology; D006056:Golgi Apparatus / Q000737:chemistry; D006624:Hippocampus / Q000201:enzymology*; D007124:Immunoenzyme Techniques; D017403:In Situ Hybridization; D008666:Metalloendopeptidases / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D015260:Neprilysin / Q000032:analysis / Q000378:metabolism; D009447:Neuroblastoma; D009457:Neuroglia / Q000201:enzymology*; D011480:Protease Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D014407:Tumor Cells, Cultured / Q000201:enzymology
D000911:Antibodies, Monoclonal; D018076:DNA, Complementary; D005904:Glial Fibrillary Acidic Protein; D006020:Glycopeptides; D011480:Protease Inhibitors; D012333:RNA, Messenger; D018826:CD13 Antigens; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D015260:Neprilysin; D000072396:Endothelin-Converting Enzymes; C008890:phosphoramidon