J. Biol. Chem., Vol. 261, Issue 33, 15519-15523, 11, 1986
Metabolism of platelet-activating factor by arachidonic acid-depleted rat polymorphonuclear leukocytes
CS Ramesha and WC Pickett
1-O-[3H]Alkyl-2-acetyl-sn-glycero-3-phosphocholine ([3H]PAF) and 1-O-
[3H]alkyl-2-lyso-sn-glycero-3-phosphocholine ([3H]lyso-PAF) when incubated
with rat polymorphonuclear leukocytes (PMN) were rapidly metabolized to
1-O-[3H]alkyl-2-acyl-sn-glycero-3-phosphocholine ([3H]alkyl-acyl-GPC)
containing long chain acyl groups in the sn-2 position. The specificity and
the absolute requirements of arachidonate (20:4) for acylation into PAF and
lyso-PAF were investigated by comparing the rate of [3H]PAF and
[3H]lyso-PAF metabolism by control rat PMN with that by rat PMN depleted of
20:4. Comparable rates of metabolism of [3H]PAF and [3H]lyso-PAF by both
control and 20:4- depleted PMN were observed at all the concentrations of
PAF and lyso- PAF studied. The nature of the fatty acyl group incorporated
into the sn-2 position of the [3H]alkyl-acyl-GPC formed was analyzed by
argentation chromatography. Dienoic fatty acids were the major fatty acid
incorporated into the alkyl-acyl-GPC by both control and 20:4- depleted PMN
at all the incubation times studied. At 3 min of incubation with [3H]PAF
and [3H]lyso-PAF, control PMN had small but significant amounts of
[3H]alkyl-acyl-GPC containing tetraenoic fatty acids, the concentration of
which gradually increased as the incubation time progressed. On the other
hand, under similar conditions, 20:4- depleted PMN had only trace amounts
of the [3H]alkyl-acyl-GPC with tetraenoic fatty acid and the concentration
of which remained at the low level throughout the incubation time. At 3 min
of incubation, the 20:4-depleted PMN had small but significant amounts of
[3H]alkyl-acyl- GPC with saturated fatty acids, the amount of which
declined by 10 min and remained at that level as the incubation time
progressed. While the concentration of [3H]alkyl-acyl-GPC with dienoic
fatty acids in the 20:4-depleted cells gradually increased with the
progress of incubation time, these molecular species of GPC in the control
PMN remained more or less constant. In spite of a very high concentration
(equivalent to that of 20:4 in control PMN) of eicosatrienoic acid (20:3
delta 5,8,11) in the 20:4-depleted PMN, no significant amounts of
[3H]alkyl-acyl-GPC with trienoic fatty acid were formed by these cells. The
rate of metabolism of [3H]PAF and [3H]lyso-PAF by the resident macrophages
isolated from control and 20:4-depleted rats was similar.(ABSTRACT
TRUNCATED AT 400 WORDS)