DIACYLGLYCEROL ELEVATIONS IN CONTROL PLATELETS ARE UNACCOMPANIED BY PLECKSTRIN PHOSPHORYLATION - IMPLICATIONS FOR THE ROLE OF DIACYLGLYCEROL IN PLATELET ACTIVATION
Mh. Fukami et H. Holmsen, DIACYLGLYCEROL ELEVATIONS IN CONTROL PLATELETS ARE UNACCOMPANIED BY PLECKSTRIN PHOSPHORYLATION - IMPLICATIONS FOR THE ROLE OF DIACYLGLYCEROL IN PLATELET ACTIVATION, European journal of biochemistry, 228(3), 1995, pp. 579-586
Several laboratories have reported that diacylglycerol levels in human
platelets (approximate to 100 pmol/10(9) platelets) increased several
fold in response to 0.5-1 U/ml thrombin. We report here fluctuations i
n diacylglycerol mass in control platelets, the magnitude of which wer
e 60-90% of that measured in platelets treated with 0.2-0.5 U/ml of th
rombin. These control platelets were not activated by such criteria as
absence of aggregation, secretion, phosphatidic acid production and p
hosphorylation of the protein kinase C substrate, pleckstrin. Thrombin
treatment evoked all of the above responses. Analysis of the diacylgl
ycerol molecular species by reverse-phase HPLC of the dimethylated, ph
osphorylated derivatives showed that all of the molecular species that
were present in control platelets were also present in thrombin-treat
ed platelets. Most of the species appeared to fluctuate at random in c
ontrol platelets with the exception of 1-stearoyl-2-arachidonoyl-sn-gl
ycerol which was more or less stable and increased severalfold over co
ntrol values only upon thrombin treatment. Furthermore, only this spec
ies accumulated as [P-32]phosphorylated PtdOH in thrombin-treated plat
elets prelabelled with [P-32]P-i. Our findings show that, in platelets
elevation of diacylglycerol molecular species other than the 1-stearo
yl-2-arachidonoyl species occurs, but these changes are not necessaril
y linked to activation of protein kinase C as measured by pleckstrin p
hosphorylation which was observed only upon elevation of 1-stearoyl-2-
arachidonoyl-sn-glycerol.