Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1994 Mar 15;298(Pt 3):669–673. doi: 10.1042/bj2980669

Purification and biochemical properties of a high-molecular-mass inositol 1,4,5-trisphosphate 3-kinase isoenzyme in human platelets.

D Communi 1, V Vanweyenberg 1, C Erneux 1
PMCID: PMC1137912  PMID: 8141781

Abstract

The phosphorylation of inositol 1,4,5-trisphosphate (InsP3) to inositol 1,3,4,5-tetrakisphosphate (InsP4) is catalysed by InsP3 3-kinase. A method is presented for a rapid purification of the enzyme from human platelets. The purified enzyme was identified as a polypeptide of M(r) 69,000-70,000 after SDS/PAGE. It had a specific activity of 1.45 +/- 0.1 mumol/min per mg, and the degree of stimulation by Ca2+/calmodulin was 17-fold at saturating calmodulin and 10 microM free Ca2+. The Km for InsP3 and for ATP was 2.0 microM and 2.5 mM respectively. Human platelet InsP3 3-kinase was not recognized by immunodetection with anti-(InsP3 3-kinase A) or anti-(InsP3 3-kinase B) antibodies. These data provide the first biochemical evidence for the existence of a novel InsP3 3-kinase isoenzyme in human platelets, which is distinct from previously reported InsP3 3-kinase A and InsP3 3-kinase B.

Full text

PDF
671

Images in this article

672Figure 5
on p.672
672Figure 6
on p.672

Selected References

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

  1. Berridge M. J., Irvine R. F. Inositol phosphates and cell signalling. Nature. 1989 Sep 21;341(6239):197–205. doi: 10.1038/341197a0. [DOI] [PubMed] [Google Scholar]
  2. Biden T. J., Altin J. G., Karjalainen A., Bygrave F. L. Stimulation of hepatic inositol 1,4,5-trisphosphate kinase activity by Ca2+-dependent and -independent mechanisms. Biochem J. 1988 Dec 15;256(3):697–701. doi: 10.1042/bj2560697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Choi K. Y., Kim H. K., Lee S. Y., Moon K. H., Sim S. S., Kim J. W., Chung H. K., Rhee S. G. Molecular cloning and expression of a complementary DNA for inositol 1,4,5-trisphosphate 3-kinase. Science. 1990 Apr 6;248(4951):64–66. doi: 10.1126/science.2157285. [DOI] [PubMed] [Google Scholar]
  4. Conigrave A., Patwardhan A., Broomhead L., Roufogalis B. A purification strategy for inositol 1,4,5-trisphosphate 3-kinase from rat liver based upon heparin interaction chromatography. Cell Signal. 1992 May;4(3):303–312. doi: 10.1016/0898-6568(92)90070-o. [DOI] [PubMed] [Google Scholar]
  5. Daniel J. L., Dangelmaier C. A., Smith J. B. Formation and metabolism of inositol 1,4,5-trisphosphate in human platelets. Biochem J. 1987 Aug 15;246(1):109–114. doi: 10.1042/bj2460109. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Erneux C., Van Sande J., Miot F., Cochaux P., Decoster C., Dumont J. E. A mechanism in the control of intracellular cAMP level: the activation of a calmodulin-sensitive phosphodiesterase by a rise of intracellular free calcium. Mol Cell Endocrinol. 1985 Dec;43(2-3):123–134. doi: 10.1016/0303-7207(85)90075-9. [DOI] [PubMed] [Google Scholar]
  7. Imboden J. B., Pattison G. Regulation of inositol 1,4,5-trisphosphate kinase activity after stimulation of human T cell antigen receptor. J Clin Invest. 1987 May;79(5):1538–1541. doi: 10.1172/JCI112986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Irvine R. F. Is inositol tetrakisphosphate the second messenger that controls Ca2+ entry into cells? Adv Second Messenger Phosphoprotein Res. 1992;26:161–185. [PubMed] [Google Scholar]
  9. King W. G., Rittenhouse S. E. Inhibition of protein kinase C by staurosporine promotes elevated accumulations of inositol trisphosphates and tetrakisphosphate in human platelets exposed to thrombin. J Biol Chem. 1989 Apr 15;264(11):6070–6074. [PubMed] [Google Scholar]
  10. Lee S. Y., Sim S. S., Kim J. W., Moon K. H., Kim J. H., Rhee S. G. Purification and properties of D-myo-inositol 1,4,5-trisphosphate 3-kinase from rat brain. Susceptibility to calpain. J Biol Chem. 1990 Jun 5;265(16):9434–9440. [PubMed] [Google Scholar]
  11. Lin A. N., Barnes S., Wallace R. W. Phosphorylation by protein kinase C inactivates an inositol 1,4,5-trisphosphate 3-kinase purified from human platelets. Biochem Biophys Res Commun. 1990 Aug 16;170(3):1371–1376. doi: 10.1016/0006-291x(90)90546-y. [DOI] [PubMed] [Google Scholar]
  12. Lin A., Wallace R. W., Barnes S. Purification and properties of a human platelet inositol 1,4,5-trisphosphate 3-kinase. Arch Biochem Biophys. 1993 Jun;303(2):412–420. doi: 10.1006/abbi.1993.1303. [DOI] [PubMed] [Google Scholar]
  13. Macphee C. H., Reifsnyder D. H., Moore T. A., Lerea K. M., Beavo J. A. Phosphorylation results in activation of a cAMP phosphodiesterase in human platelets. J Biol Chem. 1988 Jul 25;263(21):10353–10358. [PubMed] [Google Scholar]
  14. Peterson G. L. A simplification of the protein assay method of Lowry et al. which is more generally applicable. Anal Biochem. 1977 Dec;83(2):346–356. doi: 10.1016/0003-2697(77)90043-4. [DOI] [PubMed] [Google Scholar]
  15. Takazawa K., Lemos M., Delvaux A., Lejeune C., Dumont J. E., Erneux C. Rat brain inositol 1,4,5-trisphosphate 3-kinase. Ca2(+)-sensitivity, purification and antibody production. Biochem J. 1990 May 15;268(1):213–217. doi: 10.1042/bj2680213. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Takazawa K., Perret J., Dumont J. E., Erneux C. Molecular cloning and expression of a human brain inositol 1,4,5-trisphosphate 3-kinase. Biochem Biophys Res Commun. 1991 Jan 31;174(2):529–535. doi: 10.1016/0006-291x(91)91449-m. [DOI] [PubMed] [Google Scholar]
  17. Takazawa K., Perret J., Dumont J. E., Erneux C. Molecular cloning and expression of a new putative inositol 1,4,5-trisphosphate 3-kinase isoenzyme. Biochem J. 1991 Sep 15;278(Pt 3):883–886. doi: 10.1042/bj2780883. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Takazawa K., Vandekerckhove J., Dumont J. E., Erneux C. Cloning and expression in Escherichia coli of a rat brain cDNA encoding a Ca2+/calmodulin-sensitive inositol 1,4,5-trisphosphate 3-kinase. Biochem J. 1990 Nov 15;272(1):107–112. doi: 10.1042/bj2720107. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Truglia J. A., Stracher A. Purification and characterization of a calcium dependent sulfhydryl protease from human platelets. Biochem Biophys Res Commun. 1981 May 29;100(2):814–822. doi: 10.1016/s0006-291x(81)80247-1. [DOI] [PubMed] [Google Scholar]
  20. Tsujinaka T., Sakon M., Kambayashi J., Kosaki G. Cleavage of cytoskeletal proteins by two forms of Ca2+ activated neutral proteases in human platelets. Thromb Res. 1982 Oct 15;28(2):149–156. doi: 10.1016/0049-3848(82)90257-2. [DOI] [PubMed] [Google Scholar]
  21. Yamaguchi K., Hirata M., Kuriyama H. Purification and characterization of inositol 1,4,5-trisphosphate 3-kinase from pig aortic smooth muscle. Biochem J. 1988 Apr 1;251(1):129–134. doi: 10.1042/bj2510129. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES