• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of plntphysLink to Publisher's site
Plant Physiol. Dec 1988; 88(4): 1461–1468.
PMCID: PMC1055780

Adenine Nucleotide Levels, the Redox State of the NADP System, and Assimilatory Force in Nonaqueously Purified Mesophyll Chloroplasts from Maize Leaves under Different Light Intensities 1

Abstract

Recently, a nonaqueous fractionation method of obtaining highly purified mesophyll chloroplasts from maize leaves was established. This method is now used to determine adenine nucleotide levels, the redox states of the NADP system, Pi levels and dihydroxyacetone phosphate/3-phosphoglycerate ratios in mesophyll chloroplasts of Zea mays L. leaves under different light intensities. The sum of the ATP, ADP, and AMP levels was estimated to be 1.4 millimolar and the ATP/ADP ratio was 1 in the dark and 2.5 to 4 in the light. The adenine nucleotides were equilibrated by adenylate kinase. The total concentration of NADP(H) in the chloroplasts was 0.3 millimolar in the dark and 0.48 millimolar in the light. The ratio of NADPH/NADP was 0.1 to 0.18 in the dark and 0.23 to 0.48 in the light. The Pi level was estimated to be 20 millimolar in the dark and 10 to 17 millimolar in the light. The 3-phosphoglycerate reducing system was under thermodynamic equilibrium in the light. The calculated assimilatory forces were 8 per molar and 40 to 170 per molar in the dark and the light, respectively. There was no relationship between the degree of activation of pyruvate, Pi dikinase, and adenylate energy charge, or ATP/ADP ratio or ADP level under various light intensities. Only a weak relationship was found between the degree of activation of NADP-malate dehydrogenase and the NADPH/NADP ratio or NADP(H) level with increasing light intensity. A possible regulatory mechanism which is responsible for the regulation of activation of pyruvate,Pi dikinase and NADP-malate dehydrogenase is discussed.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.5M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Arnon DI. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. [PMC free article] [PubMed]
  • Ashton AR, Burnell JN, Hatch MD. Regulation of C4 photosynthesis: inactivation of pyruvate, Pi dikinase by ADP-dependent phosphorylation and activation by phosphorolysis. Arch Biochem Biophys. 1984 May 1;230(2):492–503. [PubMed]
  • Ashton AR, Hatch MD. Regulation of C4 photosynthesis: physical and kinetic properties of active (dithiol) and inactive (disulfide) NADP-malate dehydrogenase from Zea mays. Arch Biochem Biophys. 1983 Dec;227(2):406–415. [PubMed]
  • Ashton AR, Hatch MD. Regulation of C4 photosynthesis: regulation of activation and inactivation of NADP-malate dehydrogenase by NADP and NADPH. Arch Biochem Biophys. 1983 Dec;227(2):416–424. [PubMed]
  • Budde RJ, Ernst SM, Chollet R. Substrate specificity and regulation of the maize (Zea mays) leaf ADP: protein phosphotransferase catalysing phosphorylation/inactivation of pyruvate, orthophosphate dikinase. Biochem J. 1986 Jun 1;236(2):579–584. [PMC free article] [PubMed]
  • Burnell JN, Hatch MD. Regulation of C4 photosynthesis: purification and properties of the protein catalyzing ADP-mediated inactivation and Pi-mediated activation of pyruvate,Pi dikinase. Arch Biochem Biophys. 1985 Mar;237(2):490–503. [PubMed]
  • CHANCE B, BALTSCHEFFSKY H. Respiratory enzymes in oxidative phosphorylation. VII. Binding of intramitochondrial reduced pyridine nucleotide. J Biol Chem. 1958 Sep;233(3):736–739. [PubMed]
  • Foust GP, Mayhew SG, Massey V. Complex formation between ferredoxin triphosphopyridine nucleotide reductase and electron transfer proteins. J Biol Chem. 1969 Feb 10;244(3):964–970. [PubMed]
  • Gerhardt R, Heldt HW. Measurement of subcellular metabolite levels in leaves by fractionation of freeze-stopped material in nonaqueous media. Plant Physiol. 1984 Jul;75(3):542–547. [PMC free article] [PubMed]
  • Gerhardt R, Stitt M, Heldt HW. Subcellular Metabolite Levels in Spinach Leaves : Regulation of Sucrose Synthesis during Diurnal Alterations in Photosynthetic Partitioning. Plant Physiol. 1987 Feb;83(2):399–407. [PMC free article] [PubMed]
  • Hampp R, Goller M, Ziegler H. Adenylate Levels, Energy Charge, and Phosphorylation Potential during Dark-Light and Light-Dark Transition in Chloroplasts, Mitochondria, and Cytosol of Mesophyll Protoplasts from Avena sativa L. Plant Physiol. 1982 Feb;69(2):448–455. [PMC free article] [PubMed]
  • Heber UW, Santarius KA. Compartmentation and reduction of pyridine nucleotides in relation to photosynthesis. Biochim Biophys Acta. 1965 Nov 29;109(2):390–408. [PubMed]
  • Hodges TK, Leonard RT. Purification of a plasma membrane-bound adenosine triphosphatase from plant roots. Methods Enzymol. 1974;32:392–406. [PubMed]
  • Kagawa T, Bruno PL. NADP-malate dehydrogenase from leaves of Zea mays: purification and physical, chemical, and kinetic properties. Arch Biochem Biophys. 1988 Feb 1;260(2):674–695. [PubMed]
  • Kleczkowski LA, Randall DD. Maize leaf adenylate kinase : purification and partial characterization. Plant Physiol. 1986 Aug;81(4):1110–1114. [PMC free article] [PubMed]
  • Lilley RM, Chon CJ, Mosbach A, Heldt HW. The distribution of metabolites between spinach chloroplasts and medium during photosynthesis in vitro. Biochim Biophys Acta. 1977 May 11;460(2):259–272. [PubMed]
  • Nakamoto H, Edwards GE. Control of the activation/inactivation of pyruvate, Pi dikinase from the C4 plant maize by adenylate energy charge, pyruvate, and analogs of pyruvate. Biochem Biophys Res Commun. 1983 Sep 15;115(2):673–679. [PubMed]
  • Rebeille F, Hatch MD. Regulation of NADP-malate dehydrogenase in C4 plants: relationship among enzyme activity, NADPH to NADP ratios, and thioredoxin redox states in intact maize mesophyll chloroplasts. Arch Biochem Biophys. 1986 Aug 15;249(1):171–179. [PubMed]
  • Saheki S, Takeda A, Shimazu T. Assay of inorganic phosphate in the mild pH range, suitable for measurement of glycogen phosphorylase activity. Anal Biochem. 1985 Aug 1;148(2):277–281. [PubMed]
  • Stitt M, Lilley RM, Heldt HW. Adenine nucleotide levels in the cytosol, chloroplasts, and mitochondria of wheat leaf protoplasts. Plant Physiol. 1982 Oct;70(4):971–977. [PMC free article] [PubMed]
  • Usuda H. Changes in Levels of Intermediates of the C(4) Cycle and Reductive Pentose Phosphate Pathway during Induction of Photosynthesis in Maize Leaves. Plant Physiol. 1985 Aug;78(4):859–864. [PMC free article] [PubMed]
  • Usuda H. Nonaqueous purification of maize mesophyll chloroplasts. Plant Physiol. 1988 Jun;87(2):427–430. [PMC free article] [PubMed]
  • Usuda H, Edwards GE. Localization of glycerate kinase and some enzymes for sucrose synthesis in c(3) and c(4) plants. Plant Physiol. 1980 May;65(5):1017–1022. [PMC free article] [PubMed]
  • Usuda H, Ku MS, Edwards GE. Activation of NADP-Malate Dehydrogenase, Pyruvate,Pi Dikinase, and Fructose 1,6-Bisphosphatase in Relation to Photosynthetic Rate in Maize. Plant Physiol. 1984 Sep;76(1):238–243. [PMC free article] [PubMed]
  • Wirtz W, Stitt M, Heldt HW. Enzymic determination of metabolites in the subcellular compartments of spinach protoplasts. Plant Physiol. 1980 Jul;66(1):187–193. [PMC free article] [PubMed]

Articles from Plant Physiology are provided here courtesy of American Society of Plant Biologists

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...