Determination of endothelin-converting enzyme activity by high-performance liquid chromatography-on-line radioactive flow monitoring.
A rapid method to investigate the metabolism of 125I-labelled or non-labelled human big endothelin to endothelin-1 by reversed-phase high-performance liquid chromatography (HPLC) and on-line radioactive flow monitoring and/or ultraviolet detection was developed. Samples were processed by solid-phase extraction (average recovery 70-80% for non-labelled and 20-25% for 125I-labelled big endothelin and endothelin-1) followed by HPLC analysis (total analysis time 20 min). The method was successfully employed to monitor the conversion of big endothelin to endothelin-1 by various blood-borne cells, such as human polymorphonuclear leukocytes or monocytes/macrophages.
Hecker
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William Harvey Research Institute, St. Bartholomew's Hospital Medical College, Charterhouse Square, London, UK.
Kaw
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Walsh
D T
DT
D000818:Animals; D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D019332:Endothelin-1; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000378:metabolism; D006801:Humans; D007963:Leukocytes, Mononuclear / Q000378:metabolism; D008666:Metalloendopeptidases; D011498:Protein Precursors / Q000378:metabolism; D011874:Radiometry; D013056:Spectrophotometry, Ultraviolet; D013552:Swine
D019332:Endothelin-1; D016232:Endothelins; D011498:Protein Precursors; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; D000072396:Endothelin-Converting Enzymes