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. 1987 Aug 1;245(3):655–660. doi: 10.1042/bj2450655

A rapid separation method for inositol phosphates and their isomers.

K A Wreggett 1, R F Irvine 1
PMCID: PMC1148182  PMID: 3663183

Abstract

A technique is described using ACCELL QMA anion-exchange SEP-PAKs (Waters Associates) with ammonium formate-based solutions, whereby a sample can be processed within minutes to yield resolution of inositol phosphates. Isomers of inositol trisphosphate can then be separated by using this technique in combination with a rapid (5-6 min) isocratic h.p.l.c. procedure. The use of QMA SEP-PAKs offers a degree of reproducibility comparable with that of h.p.l.c. while maintaining the capacity for automation, allowing large numbers of samples to be processed rapidly.

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Selected References

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

  1. 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]
  2. 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]
  3. Berridge M. J., Downes C. P., Hanley M. R. Lithium amplifies agonist-dependent phosphatidylinositol responses in brain and salivary glands. Biochem J. 1982 Sep 15;206(3):587–595. doi: 10.1042/bj2060587. [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. 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]
  6. Clarke N. G., Dawson R. M. Alkaline O leads to N-transacylation. A new method for the quantitative deacylation of phospholipids. Biochem J. 1981 Apr 1;195(1):301–306. doi: 10.1042/bj1950301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. DAWSON R. M., HEMINGTON N., DAVENPORT J. B. Improvements in the method of determining individual phospholipids in a complex mixture by successive chemical hydrolyses. Biochem J. 1962 Sep;84:497–501. doi: 10.1042/bj0840497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. DESJOBERT A., PETEK F. Chromatographie sur papier des esters phosphoriques de l'inositol; application a l'étude de la dégradation hydrolytique de l'inositolhexaphosphate. Bull Soc Chim Biol (Paris) 1956 Sep 4;38(5-6):871–883. [PubMed] [Google Scholar]
  9. Downes C. P., Hawkins P. T., Irvine R. F. Inositol 1,3,4,5-tetrakisphosphate and not phosphatidylinositol 3,4-bisphosphate is the probable precursor of inositol 1,3,4-trisphosphate in agonist-stimulated parotid gland. Biochem J. 1986 Sep 1;238(2):501–506. doi: 10.1042/bj2380501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. HUBSCHER G., HAWTHORNE J. N. The isolation of inositol monophosphate from liver. Biochem J. 1957 Nov;67(3):523–527. doi: 10.1042/bj0670523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Hawkins P. T., Stephens L., Downes C. P. Rapid formation of inositol 1,3,4,5-tetrakisphosphate and inositol 1,3,4-trisphosphate in rat parotid glands may both result indirectly from receptor-stimulated release of inositol 1,4,5-trisphosphate from phosphatidylinositol 4,5-bisphosphate. Biochem J. 1986 Sep 1;238(2):507–516. doi: 10.1042/bj2380507. [DOI] [PMC free article] [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., Brown K. D., Berridge M. J. Specificity of inositol trisphosphate-induced calcium release from permeabilized Swiss-mouse 3T3 cells. Biochem J. 1984 Aug 15;222(1):269–272. doi: 10.1042/bj2220269. [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., Berridge M. J. Specificity of inositol phosphate-stimulated Ca2+ mobilization from Swiss-mouse 3T3 cells. Biochem J. 1986 Nov 15;240(1):301–304. doi: 10.1042/bj2400301. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Irvine R. F., Moor R. M. Micro-injection of inositol 1,3,4,5-tetrakisphosphate activates sea urchin eggs by a mechanism dependent on external Ca2+. Biochem J. 1986 Dec 15;240(3):917–920. doi: 10.1042/bj2400917. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Meek J. L. Inositol bis-, tris-, and tetrakis(phosphate)s: analysis in tissues by HPLC. Proc Natl Acad Sci U S A. 1986 Jun;83(12):4162–4166. doi: 10.1073/pnas.83.12.4162. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Richards D. E., Irvine R. F., Dawson R. M. Hydrolysis of membrane phospholipids by phospholipases of rat liver lysosomes. Biochem J. 1979 Aug 15;182(2):599–606. doi: 10.1042/bj1820599. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rouser G., Fkeischer S., Yamamoto A. Two dimensional then layer chromatographic separation of polar lipids and determination of phospholipids by phosphorus analysis of spots. Lipids. 1970 May;5(5):494–496. doi: 10.1007/BF02531316. [DOI] [PubMed] [Google Scholar]
  21. Sharps E. S., McCarl R. L. A high-performance liquid chromatographic method to measure 32P incorporation into phosphorylated metabolites in cultured cells. Anal Biochem. 1982 Aug;124(2):421–424. doi: 10.1016/0003-2697(82)90059-8. [DOI] [PubMed] [Google Scholar]
  22. Wreggett K. A., Howe L. R., Moore J. P., Irvine R. F. Extraction and recovery of inositol phosphates from tissues. Biochem J. 1987 Aug 1;245(3):933–934. doi: 10.1042/bj2450933. [DOI] [PMC free article] [PubMed] [Google Scholar]

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