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. 1987 Mar 1;242(2):361–366. doi: 10.1042/bj2420361

Separation of multiple isomers of inositol phosphates formed in GH3 cells.

N M Dean, J D Moyer
PMCID: PMC1147713  PMID: 3109388

Abstract

A high-performance-liquid-chromatography (h.p.l.c.) separation was developed, which resolves isomers of inositol monophosphate (IP), inositol bisphosphate (IP2), and inositol trisphosphate (IP3) in a single run. In GH3 cells labelled with [3H]inositol, treated with Li+ and thyrotropin-releasing hormone (TRH), radiolabelled components identified as inositol 1-phosphate (I1P), inositol 2-phosphate (I2P), inositol 4-phosphate (I4P), inositol 1,4-bisphosphate [I(1,4)P2], inositol 1,3,4-trisphosphate [I(1,3,4)P3] and inositol 1,4,5-trisphosphate [I(1,4,5)P3] are present, as are multiple unidentified IP2 peaks. After TRH stimulation, both I1P and I4P increase, the increase in I4P preceding that of I1P; I(1,4)P2 and an unknown IP2 increase; and both I(1,3,4)P3 and I(1,4,5)P3 increase, the increase in I(1,4,5)P3 being rapid and transient, whereas the increase in I(1,3,4)P3 is slower and more sustained. The most rapidly appearing inositol phosphates produced after TRH stimulation are I(1,4)P2 and I(1,4,5)P3.

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Selected References

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  1. Albert P. R., Tashjian A. H., Jr Relationship of thyrotropin-releasing hormone-induced spike and plateau phases in cytosolic free Ca2+ concentrations to hormone secretion. Selective blockade using ionomycin and nifedipine. J Biol Chem. 1984 Dec 25;259(24):15350–15363. [PubMed] [Google Scholar]
  2. Batty I. R., Nahorski S. R., Irvine R. F. Rapid formation of inositol 1,3,4,5-tetrakisphosphate following muscarinic receptor stimulation of rat cerebral cortical slices. Biochem J. 1985 Nov 15;232(1):211–215. doi: 10.1042/bj2320211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berridge M. J., Dawson R. M., Downes C. P., Heslop J. P., Irvine R. F. Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides. Biochem J. 1983 May 15;212(2):473–482. doi: 10.1042/bj2120473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berridge M. J., Irvine R. F. Inositol trisphosphate, a novel second messenger in cellular signal transduction. Nature. 1984 Nov 22;312(5992):315–321. doi: 10.1038/312315a0. [DOI] [PubMed] [Google Scholar]
  5. Binder H., Weber P. C., Siess W. Separation of inositol phosphates and glycerophosphoinositol phosphates by high-performance liquid chromatography. Anal Biochem. 1985 Jul;148(1):220–227. doi: 10.1016/0003-2697(85)90649-9. [DOI] [PubMed] [Google Scholar]
  6. Burgess G. M., McKinney J. S., Irvine R. F., Putney J. W., Jr Inositol 1,4,5-trisphosphate and inositol 1,3,4-trisphosphate formation in Ca2+-mobilizing-hormone-activated cells. Biochem J. 1985 Nov 15;232(1):237–243. doi: 10.1042/bj2320237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dawson R. M., Freinkel N., Jungalwala F. B., Clarke N. The enzymic formation of myoinositol 1:2-cyclic phosphate from phosphatidylinositol. Biochem J. 1971 May;122(4):605–607. doi: 10.1042/bj1220605. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Downes C. P., Michell R. H. The polyphosphoinositide phosphodiesterase of erythrocyte membranes. Biochem J. 1981 Jul 15;198(1):133–140. doi: 10.1042/bj1980133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Drummond A. H. Chlordiazepoxide is a competitive thyrotropin-releasing hormone receptor antagonist in GH3 pituitary tumour cells. Biochem Biophys Res Commun. 1985 Feb 28;127(1):63–70. doi: 10.1016/s0006-291x(85)80126-1. [DOI] [PubMed] [Google Scholar]
  10. Drummond A. H., Raeburn C. A. The interaction of lithium with thyrotropin-releasing hormone-stimulated lipid metabolism in GH3 pituitary tumour cells. Enhancement of stimulated 1,2-diacylglycerol formation. Biochem J. 1984 Nov 15;224(1):129–136. doi: 10.1042/bj2240129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ellis R. B., Galliard T., Hawthorne J. N. Phosphoinositides. 5. The inositol lipids of ox brain. Biochem J. 1963 Jul;88(1):125–131. doi: 10.1042/bj0880125. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. GRADO C., BALLOU C. E. Myo-inositol phosphates obtained by alkaline hydrolysis of beef brain phosphoinositide. J Biol Chem. 1961 Jan;236:54–60. [PubMed] [Google Scholar]
  13. Heslop J. P., Irvine R. F., Tashjian A. H., Jr, Berridge M. J. Inositol tetrakis- and pentakisphosphates in GH4 cells. J Exp Biol. 1985 Nov;119:395–401. doi: 10.1242/jeb.119.1.395. [DOI] [PubMed] [Google Scholar]
  14. Irvine R. F., Anggård E. E., Letcher A. J., Downes C. P. Metabolism of inositol 1,4,5-trisphosphate and inositol 1,3,4-trisphosphate in rat parotid glands. Biochem J. 1985 Jul 15;229(2):505–511. doi: 10.1042/bj2290505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Irvine R. F., Letcher A. J., Heslop J. P., Berridge M. J. The inositol tris/tetrakisphosphate pathway--demonstration of Ins(1,4,5)P3 3-kinase activity in animal tissues. Nature. 1986 Apr 17;320(6063):631–634. doi: 10.1038/320631a0. [DOI] [PubMed] [Google Scholar]
  16. Irvine R. F., Letcher A. J., Lander D. J., Downes C. P. Inositol trisphosphates in carbachol-stimulated rat parotid glands. Biochem J. 1984 Oct 1;223(1):237–243. doi: 10.1042/bj2230237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. KEMP P., HUBSCHER G., HAWTHORNE J. N. Phosphoinositides. 3. Enzymic hydrolysis of inositol-containing phospholipids. Biochem J. 1961 Apr;79:193–200. doi: 10.1042/bj0790193. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Macphee C. H., Drummond A. H. Thyrotropin-releasing hormone stimulates rapid breakdown of phosphatidylinositol 4,5-bisphosphate and phosphatidylinositol 4-phosphate in GH3 pituitary tumor cells. Mol Pharmacol. 1984 Mar;25(2):193–200. [PubMed] [Google Scholar]
  19. Michell B. Inositol phosphates. Profusion and confusion. Nature. 1986 Jan 16;319(6050):176–177. doi: 10.1038/319176a0. [DOI] [PubMed] [Google Scholar]
  20. Nishizuka Y. The role of protein kinase C in cell surface signal transduction and tumour promotion. Nature. 1984 Apr 19;308(5961):693–698. doi: 10.1038/308693a0. [DOI] [PubMed] [Google Scholar]
  21. Ramsdell J. S., Tashjian A. H., Jr Thyrotropin-releasing hormone (TRH) elevation of inositol trisphosphate and cytosolic free calcium is dependent on receptor number. Evidence for multiple rapid interactions between TRH and its receptor. J Biol Chem. 1986 Apr 25;261(12):5301–5306. [PubMed] [Google Scholar]
  22. Rebecchi M. J., Gershengorn M. C. Thyroliberin stimulates rapid hydrolysis of phosphatidylinositol 4,5-bisphosphate by a phosphodiesterase in rat mammotropic pituitary cells. Evidence for an early Ca2+-independent action. Biochem J. 1983 Nov 15;216(2):287–294. doi: 10.1042/bj2160287. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Rebecchi M. J., Monaco M. E., Gershengorn M. C. Thyrotropin releasing hormone rapidly enhances [32P]orthophosphate incorporation into phosphatidic acid in cloned GH3 cells. Biochem Biophys Res Commun. 1981 Jul 16;101(1):124–130. doi: 10.1016/s0006-291x(81)80019-8. [DOI] [PubMed] [Google Scholar]
  24. Schlegel W., Roduit C., Zahnd G. R. Polyphosphoinositide hydrolysis by phospholipase C is accelerated by thyrotropin releasing hormone (TRH) in clonal rat pituitary cells (GH3 cells). FEBS Lett. 1984 Mar 12;168(1):54–60. doi: 10.1016/0014-5793(84)80205-7. [DOI] [PubMed] [Google Scholar]
  25. Siess W. Evidence for the formation of inositol 4-monophosphate in stimulated human platelets. FEBS Lett. 1985 Jun 3;185(1):151–156. doi: 10.1016/0014-5793(85)80760-2. [DOI] [PubMed] [Google Scholar]
  26. Storey D. J., Shears S. B., Kirk C. J., Michell R. H. Stepwise enzymatic dephosphorylation of inositol 1,4,5-trisphosphate to inositol in liver. Nature. 1984 Nov 22;312(5992):374–376. doi: 10.1038/312374a0. [DOI] [PubMed] [Google Scholar]
  27. Wilson D. B., Bross T. E., Sherman W. R., Berger R. A., Majerus P. W. Inositol cyclic phosphates are produced by cleavage of phosphatidylphosphoinositols (polyphosphoinositides) with purified sheep seminal vesicle phospholipase C enzymes. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4013–4017. doi: 10.1073/pnas.82.12.4013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wilson D. B., Connolly T. M., Bross T. E., Majerus P. W., Sherman W. R., Tyler A. N., Rubin L. J., Brown J. E. Isolation and characterization of the inositol cyclic phosphate products of polyphosphoinositide cleavage by phospholipase C. Physiological effects in permeabilized platelets and Limulus photoreceptor cells. J Biol Chem. 1985 Nov 5;260(25):13496–13501. [PubMed] [Google Scholar]

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