Skip to main content
PMC home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1970 Feb;49(2):392–399. doi: 10.1172/JCI106248

Biosynthesis of cytidine diphosphate diglyceride by human platelets

Frank L Call II 1, William J Williams 1
PMCID: PMC322481  PMID: 4312547

Abstract

Homogenates of human platelets contain an enzyme which catalyzes the formation of cytidine diphosphate diglyceride from cytidine triphosphate and phosphatidic acid. The enzymatic activity could not be dissociated from platelet particles and the greatest specific activity was found in the membrane fraction. The Km for cytidine triphosphate was 0.16 mmole/liter and the apparent Km for phosphatidic acid was 6.2 mmoles/liter. The pH optimum was 7.0 and the most effective buffers were triethanolamine-HCl and Tris-HCl. The reaction was dependent on the presence of divalent cations, magnesium being the most effective of those investigated. Monovalent cations did not alter the reaction rate. Evidence is presented that the cytidine diphosphate diglyceride produced can serve as a precursor for the synthesis of phosphatidylinositol. No difference was found in the enzymatic activity in platelets from normal subjects and from patients with diseases known to interfere with platelet thromboplastic function.

Full text

PDF
392

Selected References

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

  1. ABRAMSON M. B., KATZMAN R., WILSON C. E., GREGOR H. P. IONIC PROPERTIES OF AQUEOUS DISPERSIONS OF PHOSPHATIDIC ACID. J Biol Chem. 1964 Dec;239:4066–4072. [PubMed] [Google Scholar]
  2. AGRANOFF B. W., BRADLEY R. M., BRADY R. O. The enzymatic synthesis of inositol phosphatide. J Biol Chem. 1958 Nov;233(5):1077–1083. [PubMed] [Google Scholar]
  3. ANTONIS A. The colorimetric determination of ester groups in lipid extracts. J Lipid Res. 1960 Oct;1:485–486. [PubMed] [Google Scholar]
  4. ASTER R. H., JANDL J. H. PLATELET SEQUESTRATION IN MAN. I. METHODS. J Clin Invest. 1964 May;43:843–855. doi: 10.1172/JCI104970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. BRECHER G., CRONKITE E. P. Morphology and enumeration of human blood platelets. J Appl Physiol. 1950 Dec;3(6):365–377. doi: 10.1152/jappl.1950.3.6.365. [DOI] [PubMed] [Google Scholar]
  6. Barden R. E., Cleland W. W. 1-Acylglycerol 3-phosphate acyltransferase from rat liver. J Biol Chem. 1969 Jul 10;244(13):3677–3684. [PubMed] [Google Scholar]
  7. Carter J. R., Kennedy E. P. Enzymatic synthesis of cytidine diphosphate diglyceride. J Lipid Res. 1966 Sep;7(5):678–683. [PubMed] [Google Scholar]
  8. Cohen P., Gardner F. H. Platelet preservation. IV. Preservation of human platelet concentrates by controlled slow freezing in a glycerol medium. N Engl J Med. 1966 Jun 23;274(25):1400–1407. doi: 10.1056/NEJM196606232742502. [DOI] [PubMed] [Google Scholar]
  9. DAVIDSON F. M., LONG C. The structure of the naturally occurring phosphoglycerides. 4. Action of cabbage-leaf phospholipase D on ovolecithin and related substances. Biochem J. 1958 Jul;69(3):458–466. doi: 10.1042/bj0690458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. DODGE J. T., MITCHELL C., HANAHAN D. J. The preparation and chemical characteristics of hemoglobin-free ghosts of human erythrocytes. Arch Biochem Biophys. 1963 Jan;100:119–130. doi: 10.1016/0003-9861(63)90042-0. [DOI] [PubMed] [Google Scholar]
  11. FERGUSON J. H., LEWIS J. H., ZUCKER M. B. Bleeding tendency in uremia. Blood. 1956 Dec;11(12):1073–1076. [PubMed] [Google Scholar]
  12. FIRKIN B. G., WILLIAMS W. J. The incorporation of radioactive phosphorus into the phospholipids of human leukemic leukocytes and platelets. J Clin Invest. 1961 Feb;40:423–432. doi: 10.1172/JCI104270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. GROSSMAN C. M., KOHN R. ENZYMATIC CHARACTERISTICS OF IN-VITRO INCORPORATION OF P32 ORTHOPHOSPHATE INTO HUMAN PLATELET PHOSPHATIDE. Thromb Diath Haemorrh. 1965 Mar 15;13:126–135. [PubMed] [Google Scholar]
  14. GROSSMAN C. M., KOHN R., KOCH R. Possible errors in the use of P-32 orthophosphate for the estimation of platelet life span. Blood. 1963 Jul;22:9–18. [PubMed] [Google Scholar]
  15. Grossman C. M., Bartos F. Succinate dependence of in vitro incorporation of 32P-orthophosphate into human platelet phosphatide. Arch Biochem Biophys. 1968 Oct;128(1):231–235. doi: 10.1016/0003-9861(68)90026-x. [DOI] [PubMed] [Google Scholar]
  16. HOKIN L. E., HOKIN M. R. Phosphoinositides and protein secretion in pancreas slices. J Biol Chem. 1958 Oct;233(4):805–810. [PubMed] [Google Scholar]
  17. HOKIN M. R., BENFEY B. G., HOKIN L. E. Phospholipides and adrenaline secretion in guinea pig adrenal medulla. J Biol Chem. 1958 Oct;233(4):814–817. [PubMed] [Google Scholar]
  18. HOKIN M. R., HOKIN L. E., SAFFRAN M., SCHALLY A. V., ZIMMERMANN B. U. Phospholipides and the secretion of adrenocorticotropin and of corticosteroids. J Biol Chem. 1958 Oct;233(4):811–813. [PubMed] [Google Scholar]
  19. HUBSCHER G., CLARK B. Metabolism of phospholipids. II. Isolation and properties of phosphatidic acid from mammalian liver. Biochim Biophys Acta. 1960 Jun 17;41:45–54. doi: 10.1016/0006-3002(60)90367-x. [DOI] [PubMed] [Google Scholar]
  20. Hokin M. R., Hokin L. E. The formation and continuous turnover of a fraction of phosphatidic acid on stimulation of NaC1 secretion by acetylcholine in the salt gland. J Gen Physiol. 1967 Mar;50(4):793–811. doi: 10.1085/jgp.50.4.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hutchison H. T., Cronan J. E., Jr The synthesis of cytidine diphosphate diglyceride by cell-free extracts of yeast. Biochim Biophys Acta. 1968 Dec 18;164(3):606–608. doi: 10.1016/0005-2760(68)90193-8. [DOI] [PubMed] [Google Scholar]
  22. KAI M., JOSHITA T., SAGA M. FATTY ACID COMPOSITION OF LECITHIN FROM BEEF BRAIN AND EGG YOLK. J Biochem. 1963 Nov;54:403–411. doi: 10.1093/oxfordjournals.jbchem.a127806. [DOI] [PubMed] [Google Scholar]
  23. KANFER J., KENNEDY E. P. METABOLISM AND FUNCTION OF BACTERIAL LIPIDS. II. BIOSYNTHESIS OF PHOSPHOLIPIDS IN ESCHERICHIA COLI. J Biol Chem. 1964 Jun;239:1720–1726. [PubMed] [Google Scholar]
  24. KATES M. Hydrolysis of lecithin by plant plastid enzymes. Can J Biochem Physiol. 1955 Jul;33(4):575–589. [PubMed] [Google Scholar]
  25. KIYASU J. Y., PIERINGER R. A., PAULUS H., KENNEDY E. P. The biosynthesis of phosphatidylglycerol. J Biol Chem. 1963 Jul;238:2293–2298. [PubMed] [Google Scholar]
  26. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  27. Long C., Staples D. A. Chromatographic separation of brain lipids: cerebroside and sulphatide. Biochem J. 1961 Jan;78(1):179–185. doi: 10.1042/bj0780179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. MUSHETT C. W., GOLDSMITH D. P., KELLEY K. L. Studies on lipide anticoagulants. I. Assays in vitro. J Biol Chem. 1954 Nov;211(1):163–168. [PubMed] [Google Scholar]
  29. Majerus P. W., Smith M. B., Clamon G. H. Lipid metabolism in human platelets. I. Evidence for a complete fatty acid synthesizing system. J Clin Invest. 1969 Jan;48(1):156–164. doi: 10.1172/JCI105964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Marcus A. J., Ullman H. L., Safier L. B. Lipid composition of subcellular particles of human blood platelets. J Lipid Res. 1969 Jan;10(1):108–114. [PubMed] [Google Scholar]
  31. Marcus A. J., Zucker-Franklin D., Safier L. B., Ullman H. L. Studies on human platelet granules and membranes. J Clin Invest. 1966 Jan;45(1):14–28. doi: 10.1172/JCI105318. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McCaman R. E., Finnerty W. R. Biosynthesis of cytidine diphosphate-diglyceride by a particulate fracgion from Micrococcus cerificans. J Biol Chem. 1968 Oct 10;243(19):5074–5080. [PubMed] [Google Scholar]
  33. McClure P. D., Ingram G. I., Stacey R. S., Glass U. H., Matchett M. O. Platelet function tests in thrombocythaemia and thrombocytosis. Br J Haematol. 1966 Jul;12(4):478–498. doi: 10.1111/j.1365-2141.1966.tb05656.x. [DOI] [PubMed] [Google Scholar]
  34. PAULUS H., KENNEDY E. P. The enzymatic synthesis of inositol monophosphatide. J Biol Chem. 1960 May;235:1303–1311. [PubMed] [Google Scholar]
  35. PIERINGER R. A., HOKIN L. E. Biosynthesis of phosphatidic acid from lysophosphatidic acid and palmityl coenzyme A. J Biol Chem. 1962 Mar;237:659–663. [PubMed] [Google Scholar]
  36. Perper R. J., Zee T. W., Mickelson M. M. Purification of lymphocytes and platelets by gradient centrifugation. J Lab Clin Med. 1968 Nov;72(5):842–848. [PubMed] [Google Scholar]
  37. Petzold G. L., Agranoff B. W. The biosynthesis of cytidine diphosphate diglyceride by embryonic chick brain. J Biol Chem. 1967 Mar 25;242(6):1187–1191. [PubMed] [Google Scholar]
  38. Prottey C., Hawthorne J. N. The biosynthesis of phosphatidic acid and phosphatidylinositol in mammalian pancreas. Biochem J. 1967 Oct;105(1):379–392. doi: 10.1042/bj1050379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. RABINOWITZ Y. SEPARATION OF LYMPHOCYTES, POLYMORPHONUCLEAR LEUKOCYTES AND MONOCYTES ON GLASS COLUMNS, INCLUDING TISSUE CULTURE OBSERVATIONS. Blood. 1964 Jun;23:811–828. [PubMed] [Google Scholar]
  40. RHODES D. N., LEA C. H. Phospholipids. IV. On the composition of hen's egg phospholipids. Biochem J. 1957 Mar;65(3):526–533. doi: 10.1042/bj0650526. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. ROBERTSON A. F., LANDS W. E. Positional specificites in phospholipid hydrolyses. Biochemistry. 1962 Sep;1:804–810. doi: 10.1021/bi00911a012. [DOI] [PubMed] [Google Scholar]
  42. Skipski V. P., Peterson R. F., Barclay M. Quantitative analysis of phospholipids by thin-layer chromatography. Biochem J. 1964 Feb;90(2):374–378. doi: 10.1042/bj0900374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Stanacev N. Z., Chang Y. Y., Kennedy E. P. Biosynthesis of cardiolipin in Escherichia coli. J Biol Chem. 1967 Jun 25;242(12):3018–3019. [PubMed] [Google Scholar]
  44. TATTRIE N. H., CYR R. FATTY ACID COMPOSITIONS OF NATURALLY OCCURRING LYSOLECITHINS AND LECITHINS. Biochim Biophys Acta. 1963 Dec 27;70:693–696. doi: 10.1016/0006-3002(63)90815-1. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES