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J. Biol. Chem., Vol. 263, Issue 1, 174-181, Jan, 1988
JA Maclouf and RC Murphy
Transformation of leukotriene (LT) A4 into leukotriene C4 has been found to
be carried out by human platelets in a rather efficient manner. LTC4 was
characterized by a combination of high performance liquid chromatography,
UV spectrophotometry, use of labeled precursor, guinea pig ileum bioassay,
and enzyme immunoassay. LTA4 metabolism was found to be
substrate-dependent, time-dependent, and proportional to platelet
concentration even at sub- or supraphysiological levels (0.0019-1 X 10(9)
platelets/ml). Neither plasma alone nor the supernatant of resting or
activated platelets was found to catalyze the production of LTC4 in the
presence or in the absence of reduced glutathione. These data suggest that
platelets contain a glutathione S- transferase specific for LTC4
biosynthesis. The formation of LTC4 was greatly enhanced when LTA4 was
incubated with platelets in the presence of albumin. Low concentrations of
albumin (2-4 g/liter) stabilized LTA4 to an extent that conversion into
LTC4 by the platelets could be detected after 1 h of incubation. The
possible intercellular transfer of LTA4 between neutrophils and platelets
was tested. The production of LTC4 by neutrophils was greatly enhanced in
the presence of platelets. Furthermore, the supernatant of neutrophils
stimulated with the calcium ionophore contained a short-lived acid-labile
substance which was converted by the platelets into LTC4. When platelets
were prelabeled with [35S]cysteine to allow intracellular synthesis of
[35S]glutathione, the coincubation of both cell types challenged with the
calcium ionophore resulted in the production of [35S] LTC4. These data
indicate that platelets can produce large amounts of LTC4 from
neutrophil-derived LTA4. They also suggest that such interactions may occur
in vivo and that platelets could be an important contribution to the
generation of the biologically active LTC4.
Transcellular metabolism of neutrophil-derived leukotriene A4 by human platelets. A potential cellular source of leukotriene C4
Department of Pharmacology, University of Colorado Health Sciences Center, Denver 80262.
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