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. 1973 Jul;52(7):1595–1600. doi: 10.1172/JCI107336

Identification of Inorganic Pyrophosphate in Human Platelets and Its Release on Stimulation with Thrombin

Donald C Silcox 1, Sergio Jacobelli 1, Daniel J McCarty 1
PMCID: PMC302430  PMID: 4352460

Abstract

Serum inorganic pyrophosphate (PPi) levels were consistently two- to threefold higher than plasma PPi prepared from the same blood. PPi was found in platelets in amounts ranging from 1.4 to 3 nmol/108 cells, using three different techniques for quantification. These levels are approximately 800 times higher than the mean PPi concentration in normal plasma and approximate the levels of ADP found in platelets by other workers.

About 50% of platelet PPi was specifically released extracellularly after stimulation with thrombin. Timed release experiments showed a pattern of release that resembled that described for ADP and ATP. This pattern was clearly different from that shown by platelet calcium, serotonin, or β-glucuronidase. Platelet inorganic pyrophosphatase was not released into the supernate in detectable amounts.

Platelets from patients with nucleotide storage pool deficiency showed greatly reduced levels of PPi as compared with control. There was no detectable release of PPi into extracellular medium after thrombin addition to a suspension of these platelets.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Baehner R. L., Karnovsky M. J., Karnovsky M. L. Degranulation of leukocytes in chronic granulomatous disease. J Clin Invest. 1969 Jan;48(1):187–192. doi: 10.1172/JCI105967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. CONTRACTOR S. F. THE ESTIMATION OF 5-HYDROXYTRYPTAMINE IN HUMAN BLOOD. Biochem Pharmacol. 1964 Sep;13:1351–1357. doi: 10.1016/0006-2952(64)90235-7. [DOI] [PubMed] [Google Scholar]
  3. CROSTI P. F., LUCCHELLI P. E. An easy method to determine the serotonin content of human platelets. J Clin Pathol. 1962 Mar;15:191–193. doi: 10.1136/jcp.15.2.191. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DATTA N., ZAJICEK J. Presence of pyrophosphatase in platelets. Acta Haematol. 1954 Aug;12(2):81–86. doi: 10.1159/000204604. [DOI] [PubMed] [Google Scholar]
  5. GERARDE H. W. DIRECT COUNTING OF PLATELETS. Am J Med Technol. 1964 May-Jun;30:205–207. [PubMed] [Google Scholar]
  6. HALL R. J. An improved method for the micro-determination of inorganic phosphate in small volumes of biological fluids. J Med Lab Technol. 1963 Apr;20:97–103. [PubMed] [Google Scholar]
  7. Holmsen H., Day H. J. The selectivity of the thrombin-induced platelet release reaction: subcellular localization of released and retained constituents. J Lab Clin Med. 1970 May;75(5):840–855. [PubMed] [Google Scholar]
  8. Holmsen H., Weiss H. J. Further evidence for a deficient storage pool of adenine nucleotides in platelets from some patients with thrombocytopathia--"storage pool disease". Blood. 1972 Feb;39(2):197–209. [PubMed] [Google Scholar]
  9. Holmsen H., Weiss H. J. Hereditary defect in the platelet release reaction caused by a deficiency in the storage pool of platelet adenine nucleotides. Br J Haematol. 1970 Nov;19(5):643–649. doi: 10.1111/j.1365-2141.1970.tb01648.x. [DOI] [PubMed] [Google Scholar]
  10. Ireland D. M. Effect of thrombin on the radioactive nucleotides of human washed platelets. Biochem J. 1967 Nov;105(2):857–867. doi: 10.1042/bj1050857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Johnson J. C., Shanoff M., Bass S. T., Boezi J. A., Hansen R. G. An enzymic method for determination of inorganic pyrophosphate and its use as an assay for RNA polymerase. Anal Biochem. 1968 Oct 10;26(1):137–145. doi: 10.1016/0003-2697(68)90037-7. [DOI] [PubMed] [Google Scholar]
  12. McCarty D. J., Solomon S. D., Warnock M. L., Paloyan E. Inorganic pyrophosphate concentrations in the synovial fluid of arthritic patients. J Lab Clin Med. 1971 Aug;78(2):216–229. [PubMed] [Google Scholar]
  13. Rosenberg H. The isolation and identification of "volutin" granules from Tetrahymena. Exp Cell Res. 1966 Feb;41(2):397–410. doi: 10.1016/s0014-4827(66)80147-7. [DOI] [PubMed] [Google Scholar]
  14. Russell R. G., Bisaz S., Donath A., Morgan D. B., Fleisch H. Inorganic pyrophosphate in plasma in normal persons and in patients with hypophosphatasia, osteogenesis imperfecta, and other disorders of bone. J Clin Invest. 1971 May;50(5):961–969. doi: 10.1172/JCI106589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. SUGINO Y., MIYOSHI Y. THE SPECIFIC PRECIPITATION OF ORTHOPHOSPHATE AND SOME BIOCHEMICAL APPLICATIONS. J Biol Chem. 1964 Jul;239:2360–2364. [PubMed] [Google Scholar]
  16. Solomons C. C., Styner J. Osteogenesis imperfecta: effect of magnesium administration on pyrophosphate metabolism. Calcif Tissue Res. 1969;3(4):318–326. doi: 10.1007/BF02058674. [DOI] [PubMed] [Google Scholar]
  17. Weiss H. J., Rogers J. Thrombocytopathia due to abnormalities in platelet release reaction--studies on six unrelated patients. Blood. 1972 Feb;39(2):187–196. [PubMed] [Google Scholar]

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