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. 1989 Dec 1;264(2):323–333. doi: 10.1042/bj2640323

The quantitative spectrum of inositol phosphate metabolites in avian erythrocytes, analysed by proton n.m.r. and h.p.l.c. with direct isomer detection.

T Radenberg 1, P Scholz 1, G Bergmann 1, G W Mayr 1
PMCID: PMC1133586  PMID: 2604720

Abstract

The spectrum of inositol phosphate isomers present in avian erythrocytes was investigated in qualitative and quantitative terms. Inositol phosphates were isolated in micromolar quantities from turkey blood by anion-exchange chromatography on Q-Sepharose and subjected to proton n.m.r. and h.p.l.c. analysis. We employed a h.p.l.c. technique with a novel, recently described complexometric post-column detection system, called 'metal-dye detection' [Mayr (1988) Biochem. J. 254, 585-591], which enabled us to identify non-radioactively labelled inositol phosphate isomers and to determine their masses. The results indicate that avian erythrocytes contain the same inositol phosphate isomers as mammalian cells. Denoted by the 'lowest-locant rule' [NC-IUB Recommendations (1988) Biochem. J. 258, 1-2] irrespective of true enantiomerism, these are Ins(1,4)P2, Ins(1,6)P2, Ins(1,3,4)P3, Ins(1,4,5)P3, Ins(1,3,4,5)P4, Ins(1,3,4,6)P4, Ins(1,4,5,6)P4, Ins(1,3,4,5,6)P5, and InsP6. Furthermore, we identified two inositol trisphosphate isomers hitherto not described for mammalian cells, namely Ins(1,5,6)P3 and Ins(2,4,5)P3. The possible position of these two isomers in inositol phosphate metabolism and implications resulting from absolute abundances of inositol phosphates are discussed.

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Selected References

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  1. BARTLETT G. R. Phosphorus assay in column chromatography. J Biol Chem. 1959 Mar;234(3):466–468. [PubMed] [Google Scholar]
  2. Balla T., Guillemette G., Baukal A. J., Catt K. J. Metabolism of inositol 1,3,4-trisphosphate to a new tetrakisphosphate isomer in angiotensin-stimulated adrenal glomerulosa cells. J Biol Chem. 1987 Jul 25;262(21):9952–9955. [PubMed] [Google Scholar]
  3. Bansal V. S., Inhorn R. C., Majerus P. W. The metabolism of inositol 1,3,4-trisphosphate to inositol 1,3-bisphosphate. J Biol Chem. 1987 Jul 15;262(20):9444–9447. [PubMed] [Google Scholar]
  4. Cerdan S., Hansen C. A., Johanson R., Inubushi T., Williamson J. R. Nuclear magnetic resonance spectroscopic analysis of myo-inositol phosphates including inositol 1,3,4,5-tetrakisphosphate. J Biol Chem. 1986 Nov 5;261(31):14676–14680. [PubMed] [Google Scholar]
  5. Connolly T. M., Wilson D. B., Bross T. E., Majerus P. W. Isolation and characterization of the inositol cyclic phosphate products of phosphoinositide cleavage by phospholipase C. Metabolism in cell-free extracts. J Biol Chem. 1986 Jan 5;261(1):122–126. [PubMed] [Google Scholar]
  6. Dean N. M., Moyer J. D. Metabolism of inositol bis-, tris-, tetrakis- and pentakis-phosphates in GH3 cells. Biochem J. 1988 Mar 1;250(2):493–500. doi: 10.1042/bj2500493. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Downes C. P., Berrie C. P., Hawkins P. T., Stephens L., Boyer J. L., Harden T. K. Receptor and G-protein-dependent regulation of turkey erythrocyte phosphoinositidase C. Philos Trans R Soc Lond B Biol Sci. 1988 Jul 26;320(1199):267–280. doi: 10.1098/rstb.1988.0076. [DOI] [PubMed] [Google Scholar]
  8. Hansen C. A., vom Dahl S., Huddell B., Williamson J. R. Characterization of inositol 1,3,4-trisphosphate phosphorylation in rat liver. FEBS Lett. 1988 Aug 15;236(1):53–56. doi: 10.1016/0014-5793(88)80284-9. [DOI] [PubMed] [Google Scholar]
  9. Hawkins P. T., Berrie C. P., Morris A. J., Downes C. P. Inositol 1,2-cyclic 4,5-trisphosphate is not a product of muscarinic receptor-stimulated phosphatidylinositol 4,5-bisphosphate hydrolysis in rat parotid glands. Biochem J. 1987 Apr 1;243(1):211–218. doi: 10.1042/bj2430211. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Inhorn R. C., Bansal V. S., Majerus P. W. Pathway for inositol 1,3,4-trisphosphate and 1,4-bisphosphate metabolism. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2170–2174. doi: 10.1073/pnas.84.8.2170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Irvine R. F., Letcher A. J., Lander D. J., Heslop J. P., Berridge M. J. Inositol(3,4)bisphosphate and inositol(1,3)bisphosphate in GH4 cells--evidence for complex breakdown of inositol(1,3,4)trisphosphate. Biochem Biophys Res Commun. 1987 Feb 27;143(1):353–359. doi: 10.1016/0006-291x(87)90672-3. [DOI] [PubMed] [Google Scholar]
  12. Isaacks R., Harkness D., Sampsell R., Adler J., Roth S., Kim C., Goldman P. Studies on avian erythrocyte metabolism. Inositol tetrakisphosphate: the major phosphate compound in the erythrocytes of the ostrich (Struthio camelus camelus). Eur J Biochem. 1977 Aug 1;77(3):567–574. doi: 10.1111/j.1432-1033.1977.tb11700.x. [DOI] [PubMed] [Google Scholar]
  13. Lanzetta P. A., Alvarez L. J., Reinach P. S., Candia O. A. An improved assay for nanomole amounts of inorganic phosphate. Anal Biochem. 1979 Nov 15;100(1):95–97. doi: 10.1016/0003-2697(79)90115-5. [DOI] [PubMed] [Google Scholar]
  14. Mayr G. W. A novel metal-dye detection system permits picomolar-range h.p.l.c. analysis of inositol polyphosphates from non-radioactively labelled cell or tissue specimens. Biochem J. 1988 Sep 1;254(2):585–591. doi: 10.1042/bj2540585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mayr G. W., Dietrich W. The only inositol tetrakisphosphate detectable in avian erythrocytes is the isomer lacking phosphate at position 3: a NMR study. FEBS Lett. 1987 Mar 23;213(2):278–282. doi: 10.1016/0014-5793(87)81505-3. [DOI] [PubMed] [Google Scholar]
  16. Morris A. J., Downes C. P., Harden T. K., Michell R. H. Turkey erythrocytes possess a membrane-associated inositol 1,4,5-trisphosphate 3-kinase that is activated by Ca2+ in the presence of calmodulin. Biochem J. 1987 Dec 1;248(2):489–493. doi: 10.1042/bj2480489. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Phillippy B. Q., White K. D., Johnston M. R., Tao S. H., Fox M. R. Preparation of inositol phosphates from sodium phytate by enzymatic and nonenzymatic hydrolysis. Anal Biochem. 1987 Apr;162(1):115–121. doi: 10.1016/0003-2697(87)90015-7. [DOI] [PubMed] [Google Scholar]
  18. Shears S. B., Parry J. B., Tang E. K., Irvine R. F., Michell R. H., Kirk C. J. Metabolism of D-myo-inositol 1,3,4,5-tetrakisphosphate by rat liver, including the synthesis of a novel isomer of myo-inositol tetrakisphosphate. Biochem J. 1987 Aug 15;246(1):139–147. doi: 10.1042/bj2460139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Shears S. B., Storey D. J., Morris A. J., Cubitt A. B., Parry J. B., Michell R. H., Kirk C. J. Dephosphorylation of myo-inositol 1,4,5-trisphosphate and myo-inositol 1,3,4-triphosphate. Biochem J. 1987 Mar 1;242(2):393–402. doi: 10.1042/bj2420393. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Stephens L. R., Hawkins P. T., Barker C. J., Downes C. P. Synthesis of myo-inositol 1,3,4,5,6-pentakisphosphate from inositol phosphates generated by receptor activation. Biochem J. 1988 Aug 1;253(3):721–733. doi: 10.1042/bj2530721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Stephens L., Hawkins P. T., Carter N., Chahwala S. B., Morris A. J., Whetton A. D., Downes P. C. L-myo-inositol 1,4,5,6-tetrakisphosphate is present in both mammalian and avian cells. Biochem J. 1988 Jan 1;249(1):271–282. doi: 10.1042/bj2490271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Szwergold B. S., Graham R. A., Brown T. R. Observation of inositol pentakis- and hexakis-phosphates in mammalian tissues by 31P NMR. Biochem Biophys Res Commun. 1987 Dec 31;149(3):874–881. doi: 10.1016/0006-291x(87)90489-x. [DOI] [PubMed] [Google Scholar]
  23. Tilly B. C., van Paridon P. A., Verlaan I., Wirtz K. W., de Laat S. W., Moolenaar W. H. Inositol phosphate metabolism in bradykinin-stimulated human A431 carcinoma cells. Relationship to calcium signalling. Biochem J. 1987 May 15;244(1):129–135. doi: 10.1042/bj2440129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Vallejo M., Jackson T., Lightman S., Hanley M. R. Occurrence and extracellular actions of inositol pentakis- and hexakisphosphate in mammalian brain. Nature. 1987 Dec 17;330(6149):656–658. doi: 10.1038/330656a0. [DOI] [PubMed] [Google Scholar]

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