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PMID: 9480890 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A dehydrogenase-mediated recycling system of NADPH in plant peroxisomes.

The Biochemical journal ·Vol. 330 ( Pt 2) ·1998-03-01 ·Pages 777-84

Corpas FJ, Barroso JB, Sandalio LM, Distefano S, Palma JM, Lupiáñez JA, Del Río LA

Abstract

The presence of the two NADP-dependent dehydrogenases of the pentose phosphate pathway has been investigated in plant peroxisomes from pea (Pisum sativum L.) leaves. Both enzymes, glucose-6-phosphate dehydrogenase (G6PDH; EC 1.1.1.49) and 6-phosphogluconate dehydrogenase (6PGDH; EC 1.1.1.44), were present in the matrix of leaf peroxisomes, and their kinetic properties were studied. G6PDH and 6PGDH showed a typical Michaelis-Menten kinetic saturation curve, and had specific activities of 12.4 and 29.6 mU/mg protein, respectively. The Km values of G6PDH and 6PGDH for glucose 6-phosphate and for 6-phosphogluconate were 107.3 and 10.2 microM, respectively. Dithiothreitol did not inhibit G6PDH activity. By isoelectric focusing of peroxisomal matrices, the G6PDH activity was resolved into three isoforms with isoelectric points of 5.55, 5.30 and 4.85. The isoelectric point of peroxisomal 6PGDH was 5.10. Immunoblot analyses of peroxisomal matrix with an antibody against yeast G6PDH revealed a single cross-reactive band of 56 kDa. Post-embedment, EM immunogold labelling of G6PDH confirmed that this enzyme was localized in the peroxisomal matrices, the thylakoid membrane and matrix of chloroplasts, and the cytosol. The presence of the two oxidative enzymes of the pentose phosphate pathway in plant peroxisomes implies that these organelles have the capacity to reduce NADP+ to NADPH for its re-utilization in the peroxisomal metabolism. NADPH is particularly required for the ascorbate-glutathione cycle, which has been recently demonstrated in plant peroxisomes [Jiménez, Hernández, del Río and Sevilla (1997) Plant Physiol. 114, 275-284] and represents an important antioxidant protection system against H2O2 generated in peroxisomes.

MeSH Terms
Ascorbic Acid/metabolism Glucosephosphate Dehydrogenase/metabolism Glutathione/metabolism Kinetics Microbodies/enzymology Microscopy, Electron NADP/metabolism Peas/enzymology Phosphogluconate Dehydrogenase/metabolism Plant Leaves/enzymology,ultrastructure
Chemicals
NADP Phosphogluconate Dehydrogenase phosphogluconate dehydrogenase (decarboxylating) Glucosephosphate Dehydrogenase Glutathione Ascorbic Acid
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Corpas F J
Departamento de Bioquímica, Biología Celular y Molecular de Plantas, Estación Experimental del Zaidín, CSIC, Apartado 419, E-18080 Granada, Spain.
Barroso J B
Sandalio L M
Distefano S
Palma J M
Lupiáñez J A
Del Río L A
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Article Info
Journal
The Biochemical journal
Abbr.
Biochem J
ISSN
0264-6021
DOI
10.1042/bj3300777
Published
1998-03-01
Pages
777-84
Language
English
Region
England
NLM ID
2984726R
PMCID
PMC1219205
Subset
IM
External Links
PubMed source DOI: 10.1042/bj3300777 PMC Full Text View journal details
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