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Regulation of threonine biosynthesis in barley seedlings (Hordeum vulgare L.)

Planta volume 140, pages 185–192 (1978)Cite this article

Abstract

Homoserine kinase was purified 700-fold by fractional ammonium sulfate precipitation, heat treatment, CM-Sephadex C-50 and DEAE-Sephadex A-50 ion exchange chromatography, and Sephadex G-100 gel filtration. The reaction products O-phosphohomoserine and ADP were the only compounds which caused considerable inhibition of homoserine kinase activity. Product inhibition studies showed non-competitive inhibition between ATP and O-phosphohomoserine and between homoserine and O-phosphohomoserine, and competitive inhibition between ATP and ADP. ADP showed non-competitive inhibition versus homoserine at suboptimal concentrations of ATP. At saturating concentrations of ATP no effect of ADP was observed. The homoserine kinase activity was negligible in the absence of K+ and the Km value for K+ was observed to be 4.3 mmol l−1. A non-competitive pattern was observed with respect to the substrates homoserine and ATP. Threonine synthase in the first green leaf of 6-day-old barley seedlings was partially purified 15-fold by ammonium sulfate fractionation and Sephadex G-100 gel chromatography. Threonine synthase was shown to require pyridoxal 5′-phosphate as coenzyme for optimum activity and the enzyme was strongly activated by S-adenosyl-L-methionine. The optimum pH for threonine synthase activity was 7 to 8.

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Abbreviations

PLP:

Pyridoxal 5′-phosphate

SAM:

S-adenosyl-L-methionine

HSP:

O-phosphohomoserine

References

  • Aarnes, H.: Homoserine kinase from barley seedlings. Plant Sci. Lett. 7, 187–194 (1976)

    Google Scholar 

  • Aarnes, H.: A lysine-sensitive aspartate kinase and two molecular forms of homoserine dehydrogenase from barley seedlings. Plant. Sci. Lett. 9, 137–145 (1977a)

    Google Scholar 

  • Aarnes, H.: Partial purification and characterization of methionine adenosyltransferase from pea seedlings. Plant Sci. Lett. 10, 381–390 (1977b)

    Google Scholar 

  • Aarnes, H., Rognes, S.E.: Threonine-sensitive aspartate kinase and homoserine dehydrogenase from Pisum sativum. Phytochemistry 13, 2717–2724 (1974)

    Google Scholar 

  • Bryan, J.K.: Amino acid biosynthesis and its regulation. In: Plant biochemistry 3. edn., pp. 525–560, Bonner, J., Varner, J.E., ed. New York-San Fransisco-London: Academic Press, 1976

    Google Scholar 

  • Bryan, J.K., Lissik, E.A., Matthews, B.F.: Changes in enzyme regulation during growth of maize. III Intracellular localization of homoserine dehydrogenase in chloroplasts. Plant Physiol. 59, 673–679 (1977)

    Google Scholar 

  • Burr, B., Walker, J., Truffa-Bachi, P., Cohen, G.N.: Homoserine kinase from Escherichia coli K 12. Eur. J. Biochem. 62, 519–526 (1976)

    Google Scholar 

  • Clandinin, M.T., Cossins, E.A.: Methionine biosynthesis in isolated Pisum sativum mitochondria. Phytochemistry 13, 585–591 (1974)

    Google Scholar 

  • Cleland, W.W.: The kinetics of enzyme-catalyzed reactions with two or more substrates or products. I. Nomenclature and rate equations. Biochim. Biophys. Acta 67, 104–137 (1963)

    Article  PubMed  Google Scholar 

  • Cohen, G.N.: The Regulation of Cell Metabolism. London: Holt, Rinehart & Winston 1968

    Google Scholar 

  • Datko, A.H., Giovanelli, J., Mudd, S.H.: Homocysteine biosynthesis in green plants. O-phosphorylhomoserine as the physiological substrate for cystathionine γ-synthase. J. Biol. Chem. 249, 1139–1155 (1974)

    Article  Google Scholar 

  • Ellsworth, R.K., St. Pierre, M.E.: Biosynthesis and inhibition of SAM: magnesium protoporphyrin methyltransferase of wheat. Photosynthetica 10, 291–301 (1976)

    Google Scholar 

  • Flavin, M., Slaughter, C.: Purification and properties of threonine synthase of Neurospora. J. Biol. Chem. 235, 1103–1108 (1960)

    Google Scholar 

  • Gallon, J.R., Butt, V.S.: L-tyrosine decarboxylase from barley roots. Biochem. J. 123, 5 P (1971)

    Google Scholar 

  • Gelder, B.F. van, Slater, E.C.: The extinction coefficient of cytochrome c. Biochim. Biophys. Acta 58, 593–595 (1962)

    Google Scholar 

  • Giovanelli, J., Mudd, S.H., Datko, A.H.: Homoserine esterification in green plants. Plant Physiol. 54, 725–736 (1974)

    Google Scholar 

  • Hugo, H. van, Gottschalk, G.: Purification and properties of 1-phosphofructokinase from Clostridium pasteurianum. Eur. J. Biochem. 48, 455–463 (1974)

    Google Scholar 

  • Lea, P.J., Norris, R.D.: t-RNA and aminoacyl-tRNA synthetases from higher plants. Progr. Phytochem. 4, 121–167 (1977)

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent. J. Biol. Chem. 193, 265–275 (1951)

    PubMed  Google Scholar 

  • Madison, J.T., Thompson, J.F.: Threonine synthase from higher plants: Stimulation by S-adenosylmethionine and inhibition by cysteine. Biochem. Biophys. Res. Comm. 71, 684–691 (1976)

    Google Scholar 

  • Mazelis, M., Miflin, B.J., Pratt, H.M.: A chloroplast-localized diaminopimelate decarboxylase in higher plants. FEBS-Lett. 64, 197–200 (1976)

    Google Scholar 

  • Miflin, B.J., Lea, P.J.: Amino acid metabolism. Ann. Rev. Plant Physiol. 28, 299–329 (1977)

    Google Scholar 

  • Miyajima, R., Shiio, I.: Regulation of aspartate family amino acid biosynthesis in Brevibacterium flavum. V. Properties of homoserine kinase. J. Biochem. 71, 219–226 (1972)

    Google Scholar 

  • Nigam, S.N., Ressler, C.: Biosynthesis of 2,4-diaminobutyric acid from L-[3H] homoserine and DL-[1-14C] aspartic acid in Lathyrus sylvestris W. Biochemistry 5, 3426–3431 (1966)

    Google Scholar 

  • Poulton, J.E., Butt, V.S.: Partial purification and properties of adenosine nucleosidase from leaves of spinach beet (Beta vulgaris L.). Planta 131, 179–185 (1976a)

    Google Scholar 

  • Poulton, J.E., Butt, V.S.: Purification and properties of S-adenosyl-L-homocysteine hydrolase from leaves of spinach beet. Arch. Biochem. Biophys. 172, 135–142 (1976b)

    Google Scholar 

  • Schnyder, J., Rottenberg, M., Erismann, K.H.: The synthesis of threonine and thiothreonine from O-phospho-homoserine by extracts prepared from higher plants. Biochem. Physiol. Pflanzen 167, 605–608 (1975)

    Google Scholar 

  • Shapiro, S.K., Ehninger, D.J.: Methods for the analysis and preparation of adenosylmethionine and adenosylhomocysteine. Anal. Biochem. 15, 323–333 (1966)

    Google Scholar 

  • Shewry, P.R., Miflin, B.J.: Properties and regulation of aspartate kinase from barley seedlings (Hordeum vulgare L.). Plant Physiol. 59, 69–73 (1977)

    Google Scholar 

  • Smith, L.: Spectrophotometric assay of cytochrome c oxidase. Meth. Biochem. Anal. 2, 427–434 (1955)

    Google Scholar 

  • Smith, T.A.: Recent advances in the biochemistry of plant amines. Phytochemistry 14, 865–890 (1975)

    Google Scholar 

  • Wengenmayer, H., Ebel, J., Grisebach, H.: Purification and properties of a S-adenosylmethionine: isoflavone 4′-O-methyltransferase from cell suspension cultures of Cicer arietinum L. Eur. J. Biochem. 50, 135–143 (1974)

    Google Scholar 

  • Wormser, E.H., Pardee, A.B.: Regulation of threonine biosynthesis in Escherichia coli. Arch. Biochem. Biophys. 78, 416–432 (1958)

    Google Scholar 

  • Yen, H., Gest, H.: Regulation of biosynthesis of aspartate family of amino acids in the photosynthesis bacterium Rhodopseudomonas palustris. Arch. Microbiol. 101, 187–210 (1974)

    Google Scholar 

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Affiliations

  1. Botanical Laboratory, University of Oslo, Blindern, P.O. Box 1045, Oslo 3, Norway

    Halvor Aarnes

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  1. Halvor Aarnes
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Aarnes, H. Regulation of threonine biosynthesis in barley seedlings (Hordeum vulgare L.). Planta 140, 185–192 (1978). https://doi.org/10.1007/BF00384919

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  • DOI: https://doi.org/10.1007/BF00384919

Key words

  • Homoserine kinase
  • Hordeum
  • Threonine synthese