Hydrolysis of polyphosphoinositides in human platelets.
Phospholipase C was purified 110 fold from human platelets. The activity of the enzyme was totally dependent upon Ca2+. The activity of the enzyme was markedly enhanced in the presence of arachidonic acid and was strongly inhibited by aminoglycoside antibiotics. The enzyme hydrolyzed endogenous polyphosphoinositides in addition to PI in Ca2+ dependent manner, suggesting the involvement of this enzyme in stimulus-linked rapid hydrolysis of polyphosphoinositides in platelets. The stimulation by thrombin of 32P-labelled human platelets induced about 30% decrease in 32P-TPI and about 220% increase in 32P-PA at the first 10 sec. The degree of hydrolysis of TPI was dependent upon the amount of agonist and it was not affected by the extracellular concentration of Ca2+. The changes in 32P-phospholipids in thrombin-stimulated platelets in the absence of Ca2+ were inhibited in a dose dependent manner by preincubation with relatively higher amount of quin 2 AM. The inhibition was completely overcome by an addition of CaCl2 to the suspending buffer. By such treatment in the absence of extracellular Ca2+, the intracellular Ca2+ concentration was significantly lowered below the basal level (less than 100 nM). Those observations suggest that TPI breakdown in thrombin-stimulated platelets is primary mediated by the agonist receptor coupling and requires at least the basal level of intracellular Ca2+.
Nakamura
K
K
Kambayashi
J
J
Suga
K
K
Hakata
H
H
Mori
T
T
D000634:Aminoquinolines / Q000494:pharmacology; D001792:Blood Platelets / Q000187:drug effects / Q000378:metabolism*; D002118:Calcium / Q000097:blood / Q000494:pharmacology; D006801:Humans; D006868:Hydrolysis; D066298:In Vitro Techniques; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols / Q000097:blood*; D013917:Thrombin / Q000494:pharmacology; D010738:Type C Phospholipases / Q000097:blood
D000634:Aminoquinolines; D019269:Phosphatidylinositol 4,5-Diphosphate; D018129:Phosphatidylinositol Phosphates; D010716:Phosphatidylinositols; D010738:Type C Phospholipases; D013917:Thrombin; C035739:Quin2; D002118:Calcium