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. 1988 Nov;88(3):879–886. doi: 10.1104/pp.88.3.879

Biosynthesis of Tetrapyrrole Pigment Precursors 1

Formation and Utilization of Glutamyl-tRNA for δ-Aminolevulinic Acid Synthesis by Isolated Enzyme Fractions from Chlorella Vulgaris

Yael J Avissar 1, Samuel I Beale 1
1Division of Biology and Medicine, Brown University, Providence, Rhode Island 02912
1

Supported by National Science Foundation Grant DMB85-18580.

PMCID: PMC1055677 PMID: 16666399

Abstract

The universal tetrapyrrole precursor δ-aminolevulinic acid (ALA) is formed from glutamate (Glu) in algae and higher plants. In the postulated reaction sequence, Glu-tRNA is produced by a Glu-tRNA synthetase, and the product serves as a substrate for a reduction step catalyzed by a pyridine nucleotide-requiring Glu-tRNA dehydrogenase. The reduced intermediate is then converted into ALA by a transaminase. An RNA and three enzyme fractions required for ALA formation from Glu have been isolated from soluble Chlorella extracts. The recombined fractions catalyzed ALA production from Glu or Glu-tRNA. The fraction containing the synthetase produced Glu-tRNA from Glu and tRNA in the presence of ATP and Mg2+. The isolated product of this reaction served as substrate for ALA production by the partially reconstituted enzyme system lacking the synthetase fraction and incapable of producing ALA from Glu. The production of ALA from Glu-tRNA by this partially reconstituted system did not require free Glu or ATP, and was not affected by added ATP. These results show that (a) free Glu-tRNA is an intermediate in the formation of ALA from Glu, (b) ATP is required only in the first step of the reaction sequence, and NADPH only in a later step, (c) Glu-tRNA production is the essential reaction catalyzed by one of the enzyme fractions, (d) this enzyme fraction is active in the absence of the other enzymes and is not required for activity of the others. The specific Glu-tRNA synthetase required for ALA formation has an approximate molecular weight of 73,000 ± 5,000 as determined by Sephadex G-100 gel filtration and native polyacrylamide gel electrophoresis. Other Glu-tRNA synthetases were present in the cell extracts but were ineffective in the the ALA-forming process.

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Selected References

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  1. Beale S. I., Castelfranco P. A. The Biosynthesis of delta-Aminolevulinic Acid in Higher Plants: II. Formation of C-delta-Aminolevulinic Acid from Labeled Precursors in Greening Plant Tissues. Plant Physiol. 1974 Feb;53(2):297–303. doi: 10.1104/pp.53.2.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  3. Deutscher M. P. Rat liver glutamyl ribonucleic acid synthetase. II. Further properties and anomalous pyrophosphate exchange. J Biol Chem. 1967 Mar 25;242(6):1132–1139. [PubMed] [Google Scholar]
  4. ELLIOTT W. H., COLEMAN G. A method for studying amino acid activation in crude enzyme preparations. Biochim Biophys Acta. 1962 Feb 26;57:236–244. doi: 10.1016/0006-3002(62)91116-2. [DOI] [PubMed] [Google Scholar]
  5. Ford S. H., Friedmann H. C. Formation of delta-aminolevulinic acid from glutamic acid by a partially purified enzymes system from wheat leaves. Biochim Biophys Acta. 1979 Aug 15;569(2):153–158. doi: 10.1016/0005-2744(79)90050-0. [DOI] [PubMed] [Google Scholar]
  6. Harel E., Ne'eman E. Alternative Routes for the Synthesis of 5-Aminolevulinic Acid in Maize Leaves : II. Formation from Glutamate. Plant Physiol. 1983 Aug;72(4):1062–1067. doi: 10.1104/pp.72.4.1062. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Huang D. D., Wang W. Y. Chlorophyll biosynthesis in Chlamydomonas starts with the formation of glutamyl-tRNA. J Biol Chem. 1986 Oct 15;261(29):13451–13455. [PubMed] [Google Scholar]
  8. Kern D., Lapointe J. Glutamyl transfer ribonucleic acid synthetase of Escherichia coli. Study of the interactions with its substrates. Biochemistry. 1979 Dec 25;18(26):5809–5818. doi: 10.1021/bi00593a010. [DOI] [PubMed] [Google Scholar]
  9. Kern D., Potier S., Boulanger Y., Lapointe J. The monomeric glutamyl-tRNA synthetase of Escherichia coli. Purification and relation between its structural and catalytic properties. J Biol Chem. 1979 Jan 25;254(2):518–524. [PubMed] [Google Scholar]
  10. 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]
  11. MAUZERALL D., GRANICK S. The occurrence and determination of delta-amino-levulinic acid and porphobilinogen in urine. J Biol Chem. 1956 Mar;219(1):435–446. [PubMed] [Google Scholar]
  12. Mayer S. M., Beale S. I., Weinstein J. D. Enzymatic conversion of glutamate to delta-aminolevulinic acid in soluble extracts of Euglena gracilis. J Biol Chem. 1987 Sep 15;262(26):12541–12549. [PubMed] [Google Scholar]
  13. Oh-hama T., Stolowich N. J., Scott A. I. 5-Aminolevulinic acid formation from glutamate via the C5 pathway in Clostridium thermoaceticum. FEBS Lett. 1988 Feb 8;228(1):89–93. doi: 10.1016/0014-5793(88)80591-x. [DOI] [PubMed] [Google Scholar]
  14. Proulx M., Duplain L., Lacoste L., Yaguchi M., Lapointe J. The monomeric glutamyl-tRNA synthetase from Bacillus subtilis 168 and its regulatory factor. Their purification, characterization, and the study of their interaction. J Biol Chem. 1983 Jan 25;258(2):753–759. [PubMed] [Google Scholar]
  15. Ratinaud M. H., Thomes J. C., Julien R. Glutamyl-tRNA synthetases from wheat. Isolation and characterization of three dimeric enzymes. Eur J Biochem. 1983 Oct 3;135(3):471–477. doi: 10.1111/j.1432-1033.1983.tb07675.x. [DOI] [PubMed] [Google Scholar]
  16. Renaud M., Bacha H., Remy P., Ebel J. P. Conformational activation of the yeast phenylalanyl-tRNA synthetase catalytic site induced by tRNAPhe interaction: triggering of adenosine or CpCpA trinucleoside diphosphate aminoacylation upon binding of tRNAPhe lacking these residues. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1606–1608. doi: 10.1073/pnas.78.3.1606. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rieble S., Beale S. I. Transformation of glutamate to delta-aminolevulinic acid by soluble extracts of Synechocystis sp. PCC 6803 and other oxygenic prokaryotes. J Biol Chem. 1988 Jun 25;263(18):8864–8871. [PubMed] [Google Scholar]
  18. Schneegurt M. A., Beale S. I. Characterization of the RNA Required for Biosynthesis of delta-Aminolevulinic Acid from Glutamate : Purification by Anticodon-Based Affinity Chromatography and Determination That the UUC Glutamate Anticodon Is a General Requirement for Function in ALA Biosynthesis. Plant Physiol. 1988 Feb;86(2):497–504. doi: 10.1104/pp.86.2.497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Schön A., Krupp G., Gough S., Berry-Lowe S., Kannangara C. G., Söll D. The RNA required in the first step of chlorophyll biosynthesis is a chloroplast glutamate tRNA. Nature. 1986 Jul 17;322(6076):281–284. doi: 10.1038/322281a0. [DOI] [PubMed] [Google Scholar]
  20. Thomes J. C., Ratinaud M. H., Julien R. Dimeric glutamyl-tRNA synthetases from wheat. Kinetic properties and functional structures. Eur J Biochem. 1983 Oct 3;135(3):479–484. doi: 10.1111/j.1432-1033.1983.tb07676.x. [DOI] [PubMed] [Google Scholar]
  21. URATA G., GRANICK S. Biosynthesis of alpha-aminoketones and the metabolism of aminoacetone. J Biol Chem. 1963 Feb;238:811–820. [PubMed] [Google Scholar]
  22. Wang W. Y., Huang D. D., Stachon D., Gough S. P., Kannangara C. G. Purification, Characterization, and Fractionation of the delta-Aminolevulinic Acid Synthesizing Enzymes from Light-Grown Chlamydomonas reinhardtii Cells. Plant Physiol. 1984 Mar;74(3):569–575. doi: 10.1104/pp.74.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Weinstein J. D., Beale S. I. Enzymatic conversion of glutamate to delta-aminolevulinate in soluble extracts of the unicellular green alga, Chlorella vulgaris. Arch Biochem Biophys. 1985 Mar;237(2):454–464. doi: 10.1016/0003-9861(85)90299-1. [DOI] [PubMed] [Google Scholar]
  24. Weinstein J. D., Mayer S. M., Beale S. I. Formation of delta-Aminolevulinic Acid from Glutamic Acid in Algal Extracts : Separation into an RNA and Three Required Enzyme Components by Serial Affinity Chromatography. Plant Physiol. 1987 Jun;84(2):244–250. doi: 10.1104/pp.84.2.244. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Weinstein J. D., Mayer S. M., Beale S. I. Stimulation of delta-Aminolevulinic Acid Formation in Algal Extracts by Heterologous RNA. Plant Physiol. 1986 Dec;82(4):1096–1101. doi: 10.1104/pp.82.4.1096. [DOI] [PMC free article] [PubMed] [Google Scholar]

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