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. 1987 Jul;84(3):619–623. doi: 10.1104/pp.84.3.619

Inhibition of Spinach Leaf NADPH(NADH)-Glyoxylate Reductase by Acetohydroxamate, Aminooxyacetate, and Glycidate 1

Leszek A Kleczkowski 1,2,2, Douglas D Randall 1,2, Dale G Blevins 1,2
PMCID: PMC1056639  PMID: 16665491

Abstract

Acetohydroxamate (AHA) and aminooxyacetate (AOA) were found to be potent inhibitors of purified NADPH(NADH)-dependent glyoxylate reductase from spinach (Spinacia oleracea) leaves. AHA was a noncompetitive (ro mixed) inhibitor of the NADPH-dependent activity of the reductase with a Ki of 0.33 millimolar. With NADH serving as a cofactor, AHA preferentially bound to the same form of the enzyme as glyoxylate, exhibiting a Ki of 0.31 millimolar. Glycine hydroxamate and l-glutamic acid-γ-hydroxamate were also inhibitory, but to a lesser extent than AHA. Inhibition by AOA (Ki of 1.8 millimolar) was enhanced by increased concentrations of glyoxylate, indicating that the inhibitor preferentially reacted with the glyoxylate-bound form of the enzyme. Glycidate, an effector of glycolate metabolism in leaves, was found to be a much weaker inhibitor of the enzyme with a Ki of 21 millimolar. While the inhibition by both AHA and AOA was fully reversible, glycidate acted as a tight-binding inhibitor. These findings are discussed with respect to the use of AHA, AOA, and glycidate as inhibitors of photorespiratory carbon metabolism in leaves. Caution is recommended in the use of these inhibitors with intact tissue experiments due to their lack of specificity.

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Selected References

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  1. Bergman B., Renström E., Hällbom L., Codd G. A. Effects of Aminooxyacetate and Aminoacetonitrile on Glycolate and Ammonia Release by the Cyanobacterium Anabaena cylindrica. Plant Physiol. 1985 Mar;77(3):536–539. doi: 10.1104/pp.77.3.536. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Berry J. A., Osmond C. B., Lorimer G. H. Fixation of O(2) during Photorespiration: Kinetic and Steady-State Studies of the Photorespiratory Carbon Oxidation Cycle with Intact Leaves and Isolated Chloroplasts of C(3) Plants. Plant Physiol. 1978 Dec;62(6):954–967. doi: 10.1104/pp.62.6.954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chollet R. Effect of glycidate on glycolate formation and photosynthesis in isolated spinach chloroplasts. Plant Physiol. 1976 Feb;57(2):237–240. doi: 10.1104/pp.57.2.237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Créach E., Stewart C. R. Effects of aminoacetonitrile on net photosynthesis, ribulose-1,5-bisphosphate levels, and glycolate pathway intermediates. Plant Physiol. 1982 Nov;70(5):1444–1448. doi: 10.1104/pp.70.5.1444. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Ericsson F., Carlmark B., Eliasson K. Erythrocyte and total body potassium in untreated primary hypertension. Acta Med Scand. 1981;209(6):439–444. doi: 10.1111/j.0954-6820.1981.tb11626.x. [DOI] [PubMed] [Google Scholar]
  6. FISHBEIN W. N., WINTER T. S., DAVIDSON J. D. UREASE CATALYSIS. I. STOICHIOMETRY, SPECIFICITY, AND KINETICS OF A SECOND SUBSTRATE: HYDROXYUREA. J Biol Chem. 1965 Jun;240:2402–2406. [PubMed] [Google Scholar]
  7. KOBASHI K., HASE J., UEHARA K. Specific inhibition of urease by hydroxamic acids. Biochim Biophys Acta. 1962 Dec 4;65:380–383. doi: 10.1016/0006-3002(62)91067-3. [DOI] [PubMed] [Google Scholar]
  8. Kleczkowski L. A., Randall D. D., Blevins D. G. Purification and characterization of a novel NADPH(NADH)-dependent glyoxylate reductase from spinach leaves. Comparison of immunological properties of leaf glyoxylate reductase and hydroxypyruvate reductase. Biochem J. 1986 Nov 1;239(3):653–659. doi: 10.1042/bj2390653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kohn L. D., Warren W. A. The kinetic properties of spinach leaf glyoxylic acid reductase. J Biol Chem. 1970 Aug 10;245(15):3831–3839. [PubMed] [Google Scholar]
  10. Lawyer A. L., Cornwell K. L., Gee S. L., Bassham J. A. Effects of glycine hydroxamate, carbon dioxide, and oxygen on photorespiratory carbon and nitrogen metabolism in spinach mesophyll cells. Plant Physiol. 1982 May;69(5):1136–1139. doi: 10.1104/pp.69.5.1136. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lawyer A. L., Cornwell K. L., Gee S. L., Bassham J. A. Glyoxylate and glutamate effects on photosynthetic carbon metabolism in isolated chloroplasts and mesophyll cells of spinach. Plant Physiol. 1983 Jun;72(2):420–425. doi: 10.1104/pp.72.2.420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Lawyer A. L., Zelitch I. Inhibition of glutamate:glyoxylate aminotransferase activity in tobacco leaves and callus by glycidate, an inhibitor of photorespiration. Plant Physiol. 1978 Feb;61(2):242–247. doi: 10.1104/pp.61.2.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lawyer A. L., Zelitch I. Inhibition of glycine decarboxylation and serine formation in tobacco by glycine hydroxamate and its effect on photorespiratory carbon flow. Plant Physiol. 1979 Nov;64(5):706–711. doi: 10.1104/pp.64.5.706. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mulligan R. M., Wilson B., Tolbert N. E. Effects of glyoxylate on photosynthesis by intact chloroplasts. Plant Physiol. 1983 Jun;72(2):415–419. doi: 10.1104/pp.72.2.415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Peterson R. B. Enhanced Incorporation of Tritium into Glycolate during Photosynthesis by Tobacco Leaf Tissue in the Presence of Tritiated Water. Plant Physiol. 1982 Jan;69(1):192–197. doi: 10.1104/pp.69.1.192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Randall D. D., Tolbert N. E., Gremel D. 3-Phosphoglycerate Phosphatase in Plants: II. Distribution, Physiological Considerations, and Comparison with P-Glycolate Phosphatase. Plant Physiol. 1971 Oct;48(4):480–487. doi: 10.1104/pp.48.4.480. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sarojini G., Oliver D. J. Inhibition of glycine oxidation by carboxymethoxylamine, methoxylamine, and acethydrazide. Plant Physiol. 1985 Mar;77(3):786–789. doi: 10.1104/pp.77.3.786. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Thompson C. M., Whittingham C. P. Intracellular localisation of phosphoglycollate phosphatase and glyoxalate reductase. Biochim Biophys Acta. 1967;143(3):642–644. doi: 10.1016/0005-2728(67)90074-6. [DOI] [PubMed] [Google Scholar]
  19. Tolbert N. E., Harrison M., Selph N. Aminooxyacetate stimulation of glycolate formation and excretion by chlamydomonas. Plant Physiol. 1983 Aug;72(4):1075–1083. doi: 10.1104/pp.72.4.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Tolbert N. E., Yamazaki R. K., Oeser A. Localization and properties of hydroxypyruvate and glyoxylate reductases in spinach leaf particles. J Biol Chem. 1970 Oct 10;245(19):5129–5136. [PubMed] [Google Scholar]
  21. Winkler R. G., Blevins D. G., Polacco J. C., Randall D. D. Ureide Catabolism of Soybeans : II. Pathway of Catabolism in Intact Leaf Tissue. Plant Physiol. 1987 Mar;83(3):585–591. doi: 10.1104/pp.83.3.585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. ZELITCH I., GOTTO A. M. Properties of a new glyoxylate reductase from leaves. Biochem J. 1962 Sep;84:541–546. doi: 10.1042/bj0840541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Zelitch I. Effect of glycidate, an inhibitor of glycolate synthesis in leaves, on the activity of some enzymes of the glycolate pathway. Plant Physiol. 1978 Feb;61(2):236–241. doi: 10.1104/pp.61.2.236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Zelitch I. The effect of glycidate, an inhibitor of glycolate synthesis, on photorespiration and net photosynthesis. Arch Biochem Biophys. 1974 Jul;163(1):367–377. doi: 10.1016/0003-9861(74)90488-3. [DOI] [PubMed] [Google Scholar]

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