Purification and characterisation of leukotriene A4 hydrolase from rat neutrophils |
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Authors: | JF Evans P Dupuis AW Ford-Hutchinson |
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Institution: | Merck Frosst Canada Inc., P.O. Box 1005, Pointe Claire-Dorval, Québec, H9R 4P8 Canada |
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Abstract: | Leukotriene A4 hydrolase was rapidly and extensively purified from rat neutrophils using anion exchange and gel filtration high-pressure liquid chromatography. The enzyme which converts the allylic epoxide leukotriene A4 to the 5,12-dihydroxyeicosatetraenoic acid leukotriene B4 was localized in the cytosolic fraction and exhibited an optimum activity at pH 7.8 and apparent Km for leukotriene A4 between 2 · 10?5 and 3 · 10?5 M. The purified leukotriene A4 hydrolase was shown to have a molecular weight of 68 000 on sodium dodecylsulfate polyacrylamide gel electrophoresis and of 50 000 by gel filtration. The molecular weight and monomeric native form of this enzyme are unique characteristics which distinguish leukotriene A4 hydrolase from previously purified epoxide hydrolases. |
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Keywords: | Leukotriene synthesis HPLC (Rat neutrophil) Hepes 4-(2-hydroxyethyl)-1-piperazineethane-sulfonic acid PMSF phenylmethylsulphonyl fluoride |
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