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. 1997 Oct;115(2):727–736. doi: 10.1104/pp.115.2.727

L-myo-lnositol 1-Phosphate Synthase from Plant Sources (Characteristics of the Chloroplastic and Cytosolic Enzymes).

A RayChaudhuri 1, N C Hait 1, S Dasgupta 1, T J Bhaduri 1, R Deb 1, A L Majumder 1
PMCID: PMC158533  PMID: 12223840

Abstract

L-myo-inositol 1-phosphate synthase (EC 5.5.1.4) from cyanobacterial (Spirulina platensis), algal (Euglena gracilis), and higher plant (Oryza sativa, Vigna radiata) sources was purified to electrophoretic homogeneity, biochemically characterized, and compared. Both chloroplastic and cytosolic forms of the enzyme were detected in E. gracilis, O. sativa, and V. radiata, whereas only the cytosolic form was detected in streptomycin-bleached or chloroplastic mutants of E. gracilis and in S. platensis. Both the chloroplastic and cytosolic forms from different sources could be purified following the same three-step chromatographic protocol. L-myo-inositol 1-phosphate synthases purified from these different sources do not differ significantly with respect to biochemical and kinetic parameters except for the molecular mass of the chloroplastic and cytosolic native holoenzymes, which appear to be homotetrameric and homotrimeric associations of their constituent subunits, respectively. Monovalent and divalent cations, sugar alcohols, and sugar phosphates are inhibitory to the enzyme activity. N-ethylmaleimide inhibition of synthase activity could be protected by the combined presence of the substrate glucose-6-phosphate and cofactor NAD+. Antibody raised against the cytosolic enzyme from E. gracilis immunoprecipitates and cross-reacts with both chloroplastic and cytosolic forms from the other sources studied.

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Selected References

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  1. Abu-Abied M., Holland D. The gene c-ino1 from Citrus paradisi is highly homologous to tur1 and ino1 from yeast and Spirodela encoding for myo-inositol phosphate synthase. Plant Physiol. 1994 Dec;106(4):1689–1689. doi: 10.1104/pp.106.4.1689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Adhikari J., Majumder A. L., Bhaduri T. J., Dasgupta S., Majumder A. L. Chloroplast as a Locale of L-myo-Inositol-1-Phosphate Synthase. Plant Physiol. 1987 Nov;85(3):611–614. doi: 10.1104/pp.85.3.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barnett J. E., Rasheed A., Corina D. L. Partial reactions of D-glucose 6-phosphate-1L-myoinositiol 1-phosphate cyclase. Biochem J. 1973 Jan;131(1):21–30. doi: 10.1042/bj1310021. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Biffen M., Hanke D. E. Reduction in the level of intracellular myo-inositol in cultured soybean (Glycine max) cells inhibits cell division. Biochem J. 1990 Feb 1;265(3):809–814. doi: 10.1042/bj2650809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Biswas B. B., Ghosh B., Majumder A. L. myo-Inositol polyphosphates and their role in cellular metabolism. A proposed cycle involving glucose-6-phosphate and myo-inositol phosphates. Subcell Biochem. 1984;10:237–280. doi: 10.1007/978-1-4613-2709-7_4. [DOI] [PubMed] [Google Scholar]
  6. Bohnert H. J., Nelson D. E., Jensen R. G. Adaptations to Environmental Stresses. Plant Cell. 1995 Jul;7(7):1099–1111. doi: 10.1105/tpc.7.7.1099. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Burnette W. N. "Western blotting": electrophoretic transfer of proteins from sodium dodecyl sulfate--polyacrylamide gels to unmodified nitrocellulose and radiographic detection with antibody and radioiodinated protein A. Anal Biochem. 1981 Apr;112(2):195–203. doi: 10.1016/0003-2697(81)90281-5. [DOI] [PubMed] [Google Scholar]
  9. CHEN I. W., CHARALAMPOUS F. C. BIOCHEMICAL STUDIES ON INOSITOL. VII. BIOSYNTHESIS OF INOSITOL BY A SOLUBLE ENZYME SYSTEM. J Biol Chem. 1964 Jun;239:1905–1910. [PubMed] [Google Scholar]
  10. Chen C. H., Eisenberg F., Jr Myoinosose-2 1-phosphate: an intermediate in the myoinositol 1-phosphate synthase reaction. J Biol Chem. 1975 Apr 25;250(8):2963–2967. [PubMed] [Google Scholar]
  11. Dean-Johnson M., Henry S. A. Biosynthesis of inositol in yeast. Primary structure of myo-inositol-1-phosphate synthase (EC 5.5.1.4) and functional analysis of its structural gene, the INO1 locus. J Biol Chem. 1989 Jan 15;264(2):1274–1283. [PubMed] [Google Scholar]
  12. Donahue T. F., Henry S. A. myo-Inositol-1-phosphate synthase. Characteristics of the enzyme and identification of its structural gene in yeast. J Biol Chem. 1981 Jul 10;256(13):7077–7085. [PubMed] [Google Scholar]
  13. Drøbak B. K. The plant phosphoinositide system. Biochem J. 1992 Dec 15;288(Pt 3):697–712. doi: 10.1042/bj2880697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Eisenberg F., Jr D-myoinositol 1-phosphate as product of cyclization of glucose 6-phosphate and substrate for a specific phosphatase in rat testis. J Biol Chem. 1967 Apr 10;242(7):1375–1382. [PubMed] [Google Scholar]
  15. Escamilla J. E., Contreras M., Martínez A., Zentella-Piña M. L-myoinositol-1-phosphate synthase from Neurospora crassa: purification to homogeneity and partial characterization. Arch Biochem Biophys. 1982 Oct 1;218(1):275–285. doi: 10.1016/0003-9861(82)90346-0. [DOI] [PubMed] [Google Scholar]
  16. Funkhouser E. A., Loewus F. A. Purification of myo-Inositol 1-Phosphate Synthase from Rice Cell Culture by Affinity Chromatography. Plant Physiol. 1975 Dec;56(6):786–790. doi: 10.1104/pp.56.6.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gumber S. C., Loewus M. W., Loewus F. A. myo-Inositol-1-phosphate synthase from pine pollen: sulfhydryl involvement at the active site. Arch Biochem Biophys. 1984 Jun;231(2):372–377. doi: 10.1016/0003-9861(84)90400-4. [DOI] [PubMed] [Google Scholar]
  18. Henry S. A., Atkinson K. D., Kolat A. I., Culbertson M. R. Growth and metabolism of inositol-starved Saccharomyces cerevisiae. J Bacteriol. 1977 Apr;130(1):472–484. doi: 10.1128/jb.130.1.472-484.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Henry S. A., Klig L. S., Loewy B. S. The genetic regulation and coordination of biosynthetic pathways in yeast: amino acid and phospholipid synthesis. Annu Rev Genet. 1984;18:207–231. doi: 10.1146/annurev.ge.18.120184.001231. [DOI] [PubMed] [Google Scholar]
  20. Ishitani M., Majumder A. L., Bornhouser A., Michalowski C. B., Jensen R. G., Bohnert H. J. Coordinate transcriptional induction of myo-inositol metabolism during environmental stress. Plant J. 1996 Apr;9(4):537–548. doi: 10.1046/j.1365-313x.1996.09040537.x. [DOI] [PubMed] [Google Scholar]
  21. Johnson M. D., Sussex I. M. 1 L-myo-Inositol 1-Phosphate Synthase from Arabidopsis thaliana. Plant Physiol. 1995 Feb;107(2):613–619. doi: 10.1104/pp.107.2.613. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Johnson M. D., Wang X. Differentially expressed forms of 1-L-myo-inositol-1-phosphate synthase (EC 5.5.1.4) in Phaseolus vulgaris. J Biol Chem. 1996 Jul 19;271(29):17215–17218. doi: 10.1074/jbc.271.29.17215. [DOI] [PubMed] [Google Scholar]
  23. Klig L. S., Henry S. A. Isolation of the yeast INO1 gene: located on an autonomously replicating plasmid, the gene is fully regulated. Proc Natl Acad Sci U S A. 1984 Jun;81(12):3816–3820. doi: 10.1073/pnas.81.12.3816. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Klig L. S., Zobel P. A., Devry C. G., Losberger C. Comparison of INO1 gene sequences and products in Candida albicans and Saccharomyces cerevisiae. Yeast. 1994 Jun;10(6):789–800. doi: 10.1002/yea.320100609. [DOI] [PubMed] [Google Scholar]
  25. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  26. Loewus M. W., Bedgar D. L., Loewus F. A. 1L-myo-inositol 1-phosphate synthase from pollen of Lilium longiflorum. An ordered sequential mechanism. J Biol Chem. 1984 Jun 25;259(12):7644–7647. [PubMed] [Google Scholar]
  27. Maeda T., Eisenberg F., Jr Purification, structure, and catalytic properties of L-myo-inositol-1-phosphate synthase from rat testis. J Biol Chem. 1980 Sep 25;255(18):8458–8464. [PubMed] [Google Scholar]
  28. Majumder A. L., Duttagupta S., Goldwasser P., Donahue T. F., Henry S. A. The mechanism of interallelic complementation at the INO1 locus in yeast: immunological analysis of mutants. Mol Gen Genet. 1981;184(3):347–354. doi: 10.1007/BF00352503. [DOI] [PubMed] [Google Scholar]
  29. Mauck L. A., Wong Y. H., Sherman W. R. L-myo-Inositol-1-phosphate synthase from bovine testis: purification to homogeneity and partial characterization. Biochemistry. 1980 Jul 22;19(15):3623–3629. doi: 10.1021/bi00556a031. [DOI] [PubMed] [Google Scholar]
  30. Naccarato W. F., Ray R. E., Wells W. W. Biosynthesis of myo-inositol in rat mammary gland. Isolation and properties of the enzymes. Arch Biochem Biophys. 1974 Sep;164(1):194–201. doi: 10.1016/0003-9861(74)90022-8. [DOI] [PubMed] [Google Scholar]
  31. Nikoloff D. M., Henry S. A. Genetic analysis of yeast phospholipid biosynthesis. Annu Rev Genet. 1991;25:559–583. doi: 10.1146/annurev.ge.25.120191.003015. [DOI] [PubMed] [Google Scholar]
  32. Pittner F., Fried W., Hoffmann-Ostenhof O. Studies on the biosynthesis of cyclitols, XXX. Purification of myo-inositol-1-phosphate synthase of rat testes to homogeneity by affinity chromatography on NAD-sepharose. Hoppe Seylers Z Physiol Chem. 1974 Feb;355(2):222–224. [PubMed] [Google Scholar]
  33. Smart C. C., Fleming A. J. A plant gene with homology to D-myo-inositol-3-phosphate synthase is rapidly and spatially up-regulated during an abscisic-acid-induced morphogenic response in Spirodela polyrrhiza. Plant J. 1993 Aug;4(2):279–293. doi: 10.1046/j.1365-313x.1993.04020279.x. [DOI] [PubMed] [Google Scholar]
  34. Vernon D. M., Bohnert H. J. A novel methyl transferase induced by osmotic stress in the facultative halophyte Mesembryanthemum crystallinum. EMBO J. 1992 Jun;11(6):2077–2085. doi: 10.1002/j.1460-2075.1992.tb05266.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. White M. J., Lopes J. M., Henry S. A. Inositol metabolism in yeasts. Adv Microb Physiol. 1991;32:1–51. doi: 10.1016/s0065-2911(08)60004-1. [DOI] [PubMed] [Google Scholar]
  36. Zsindely A., Szabolcs M., Aradi J., Schablik M., Kiss A., Szabó G. Investigations on myo-inositol-1-phosphate synthase from the wild type and the inositol-dependent mutant of Neurospora crassa. Acta Biol Acad Sci Hung. 1977;28(3):281–290. [PubMed] [Google Scholar]

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