Role of Reduced Glutathione on Platelet Functions

Summary

The role of reduced glutathione (GSH) on platelet functions was investigated utilizing thiol oxidizing agent, “diamide”. Diamide reacted rapidly with GSH in platelets, but not with protein thiols. Platelets treated with diamide showed inhibition of platelet aggregation induced by ADP, epinephrine and collagen in a concentration dependent manner. Clot retraction was grossly inhibited by diamide. Platelet interaction with polymerizing fibrin was examined by the method of Niewiarowski et al. (1972). There was no interaction observed between diamide-treated platelets and polymerizing fibrin. Ultrastructural observation of clots formed in the presence of diamide also showed no direct contact between platelets and fibrin strands. Platelets retained their granular contents, but showed loss of microtubules and dilatation of open canalicular system. Our findings may further support the idea that GSH plays an important role on platelet functions.

• References

• 1 Ando Y, Steiner M, Baldini MG. 1974; Effect of storage on sulfhydryl and disulfide groups of human platelets. Blood 43: 121
• 2 Barber AJ, Jamieson GA. 1970; Isolation and characterization of plasma membranes from human blood platelets. J. Biol. Chem. 245: 6357
• 3 Berlin RD, Oliver JM. 1976; Inhibition of microtubule - membrane interactions by diamide. Fed. Proc 476
• 4 Beutler E, Duron O, Kelly BM. 1963; Improved method for determination of blood glutathione. J. Lab. Chin. Med. 61: 882
• 5 Beutler E. 1972; Glanzmann’s thrombasthenia and reduced glutathione. New Engl. J. Med 287: 1094
• 6 Born GV R. 1962; Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature 194: 921
• 7 Carson PE, Fischer H. 1966; Glucose-6-phosphate dehydrogenase deficiency and related disorders of the pentose phosphate pathway. Amer. J. Med. 41: 744
• 8 Edelman GM, Yahara I, Wangm JL. 1973; Receptor mobility and receptor-cytoplasmic interactions in lymphocytes. Pro Natl. Acad. Sci. U.S.A. 70: 1442
• 9 Fairbanks G, Steck TL, Wallach FH. 1971; Electrophoretic analysis of the major polypeptides of the human erythrocyte membrane. Biochemistry 10: 2606
• 10 Gross RT, Hurwits RE, Marks PA. 1958; A hereditary enzymatic defect in erythrocyte metabolism: Glucose-6-phosphate dehydrogenase deficiency. J. Clin. Invest 37: 1176
• 11 Ikeda Y, Steiner M. 1978; Sulfhydriles of platelet tubulins; A role in polymerization and cholchicine binding activity. Biochemistry 17: 3454
• 12 Karpatokin S, Weiss HJ. 1972; Deficiency of glutathione peroxidase with hight levels of reduced glutathione in Glanzmann’s thrombasthenia. New Engl. J. Med. 287: 1062
• 13 Kosower NS, Kosower EM, Wertheim B. 1969; Diamide, a new reagent for intracellular oxidation of glutathione to the disulfide. Biochem. Biophys. Res. Comm. 37: 593
• 14 Kosower EM. 1970; A role for glutathione in muscle contraction. Experimenta 26: 760
• 15 Kosower NS, Marikovsky Y, Wertheim B, Danon D. 1971; Glutathione oxidation and biophysical aspects of injury to human erythorocytes. J. Lab. Clin. Med. 78: 533
• 16 Kosower EM, Correa W, Kinon BJ, Kosower NS. 1972; Glutathione VII. Differentiation among substrates by thiol-oxidizing agent, diamide. Bioch. Biophys. Acta 264: 39
• 17 Maizel JV. 1969. Acrylamide gel electrophoresis of proteins and nucleic acid. In Habel K, Salzman ND. (eds.) Fundamental Techiniques in Virology. Academic Press; New York: p 334
• 18 Mills DC B, Roft IA, Roberts GC K. 1968; The release of nucleotides, 5- hydroxytryptamine and enzyme from human blood platelets during aggregation. J. Physiol. 195: 715-729
• 19 Niewiarowski S, Regoeczi E, Stewart GJ, Senyi AF, Mustard JF. 1972; Platelet interaction with polymerizing fibrin. J. Chin. Invest. 51: 685-700
• 20 Nicholson GL. 1975; Transmembrane control of the receptors or normal and tumor cells. Biochim. Biophys. Acta 457: 57-108