Subcellular Localization of the Starch Degradative and Biosynthetic Enzymes of Spinach Leaves

Overview

Journal Plant Physiol

Specialty Physiology

Date 1979 Aug 1

PMID 16660929

Citations 58

Authors

T W Okita

E Greenberg

D N Kuhn

J Preiss

Affiliations

Soon will be listed here.

Abstract

The subcellular localization of the starch biosynthetic and degradative enzymes of spinach leaves was carried out by measuring the distribution of the enzymes in a crude chloroplast pellet and soluble protein fraction, and by the separation on sucrose density gradients of intact organelles, chloroplasts, peroxisomes, and mitochondria of a protoplast lysate. ADP-Glucose pyrophosphorylase, starch synthase, and starch-branching enzymes are quantitatively associated with the chloroplasts. The starch degradative enzymes amylase, R-enzyme (debranching activity), phosphorylase, and D-enzyme (transglycosylase) are observed both in the chloroplast and soluble protein fractions, the bulk of the degradative enzyme activities reside in the latter fraction. Chromatography of a chloroplast extract on diethylaminoethyl-cellulose resolves the R- and D-enzymes from amylase and phosphorylase activities although the two latter enzyme activities coeluted. The digestion pattern of amylase with amylopectin as a substrate indicates an endolytic activity but displays properties unlike the typical alpha-amylase as isolated from endosperm tissue.

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References

Nishimura M, Graham D, Akazawa T . Isolation of intact chloroplasts and other cell organelles from spinach leaf protoplasts. Plant Physiol. 1976; 58(3):309-14. PMC: 542237. DOI: 10.1104/pp.58.3.309. View

Levi C, Gibbs M . Starch degradation in isolated spinach chloroplasts. Plant Physiol. 1976; 57(6):933-5. PMC: 542152. DOI: 10.1104/pp.57.6.933. View

Pongratz P, Beck E . Diurnal oscillation of amylolytic activity in spinach chloroplasts. Plant Physiol. 1978; 62(5):687-9. PMC: 1092199. DOI: 10.1104/pp.62.5.687. View

Stitt M, Bulpin P, Ap Rees T . Pathway of starch breakdown in photosynthetic tissues of Pisum sativum. Biochim Biophys Acta. 1978; 544(1):200-14. DOI: 10.1016/0304-4165(78)90223-4. View

Wolosiuk R, Buchanan B . Activation of Chloroplast NADP-linked Glyceraldehyde-3-Phosphate Dehydrogenase by the Ferredoxin/Thioredoxin System. Plant Physiol. 1978; 61(4):669-71. PMC: 1091941. DOI: 10.1104/pp.61.4.669. View

Bahr J, Jensen R . Activation of ribulose bisphosphate carboxylase in intact chloroplasts by CO2 and light. Arch Biochem Biophys. 1978; 185(1):39-48. DOI: 10.1016/0003-9861(78)90141-8. View

Hawker J, Ozbun J, Ozaki H, Greenberg E, Preiss J . Interaction of spinach leaf adenosine diphosphate glucose alpha-1,4-glucan alpha-4-glucosyl transferase and alpha-1,4-glucan, alpha-1,4-glucan-6-glycosyl transferase in synthesis of branched alpha-glucan. Arch Biochem Biophys. 1974; 160(2):530-51. DOI: 10.1016/0003-9861(74)90430-5. View

Heldt H, Rapley L . Specific transport of inorganic phosphate, 3-phosphoglycerate and dihydroxyacetonephosphate, and of dicarboxylates across the inner membrane of spinach chloroplasts. FEBS Lett. 1970; 10(3):143-148. DOI: 10.1016/0014-5793(70)80438-0. View

Schafer G, Heber U . Glucose transport into spinach chloroplasts. Plant Physiol. 1977; 60(2):286-9. PMC: 542597. DOI: 10.1104/pp.60.2.286. View

10.

Evans R, Manners D . Studies on carbohydrate-metabolising enzymes. XXVII. A comparison of the action of yeast isoamylase and bacterial pullulanase on amylopectin. Carbohydr Res. 1971; 20(2):339-49. DOI: 10.1016/s0008-6215(00)81388-9. View

11.

Peavey D, Steup M, Gibbs M . Characterization of starch breakdown in the intact spinach chloroplast. Plant Physiol. 1977; 60(2):305-8. PMC: 542601. DOI: 10.1104/pp.60.2.305. View

12.

GHOSH H, Preiss J . Adenosine diphosphate glucose pyrophosphorylase. A regulatory enzyme in the biosynthesis of starch in spinach leaf chloroplasts. J Biol Chem. 1966; 241(19):4491-504. View

13.

Heldt H, Chon C, Maronde D . Role of orthophosphate and other factors in the regulation of starch formation in leaves and isolated chloroplasts. Plant Physiol. 1977; 59(6):1146-55. PMC: 542524. DOI: 10.1104/pp.59.6.1146. View

14.

TREVELYAN W, PROCTER D, Harrison J . Detection of sugars on paper chromatograms. Nature. 1950; 166(4219):444-5. DOI: 10.1038/166444b0. View

15.

Steup M, Peavey D, Gibbs M . The regulation of starch metabolism by inorganic phosphate. Biochem Biophys Res Commun. 1976; 72(4):1554-61. DOI: 10.1016/s0006-291x(76)80191-x. View

16.

Levi C, Preiss J . Amylopectin degradation in pea chloroplast extracts. Plant Physiol. 1978; 61(2):218-20. PMC: 1091835. DOI: 10.1104/pp.61.2.218. View

17.

Anderson L, Ng T, Park K . Inactivation of pea leaf chloroplastic and cytoplasmic glucose 6-phosphate dehydrogenases by light and dithiothreitol. Plant Physiol. 1974; 53(6):835-9. PMC: 541458. DOI: 10.1104/pp.53.6.835. View

18.

Lee E, Marshall J, WHELAN W . The substrate specificity of amylopectin-debranching enzymes from sweet corn. Arch Biochem Biophys. 1971; 143(2):365-74. DOI: 10.1016/0003-9861(71)90223-2. View

19.

Heber U, Santarius K . Direct and indirect transfer of ATP and ADP across the chloroplast envelope. Z Naturforsch B. 1970; 25(7):718-28. DOI: 10.1515/znb-1970-0714. View

20.

Arnon D . COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949; 24(1):1-15. PMC: 437905. DOI: 10.1104/pp.24.1.1. View