Abstract
Lipoxygenase activities were estimated in platelet subcellular fractions as well as in intact platelets obtained from normal subjects and 7 patients with platelet-lipoxygenase deficiency. From a washed platelet suspension (intact platelets), 12, 000g supernatant of sonicated platelets (F-I), cytosol fractions (F-II) and microsomal fractions (F-III) were prepared by differential centrifugation. 12-Hydroxyeicosatetraenoic acid (12-HETE) produced by the incubation of arachidonic acid with platelets or each fraction from them was measured by reversed-phase high-performance liquid chromatography analysis.In normal subjects, 12-HETE production by subcellular fractions was 87.1% (F-I), 31.4% (F-II) and 17.2% (F-III) of that by intact platelets, and was not significantly affected by the addition of CaCl2. Of the 7 patients, one patient showed no detectable lipoxygenase activity in any subcellular fractions and in intact platelets; the other patients showed reduced activities in all subcellular fractions as well as in intact platelets. In some of these patients, it appeared that lipoxygenase activities were not fully expressed in intact platelets because much more amounts of 12-HETE was produced by their subcellular fractions than by intact platelets especially in the presence of CaCl2.
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