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Subcellular distribution of multiple forms of glutathione reductase in leaves of pea (Pisum sativum L.)

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Abstract

On sodium-dodecyl-sulfate polyacrylamide gels, purified glutathione reductase (GR; EC 1.6.4.2) from the leaves of two- to three-week-old pea (Pisum sativum L. cv. Birte) seedlings was represented by a single band with an apparent molecular weight of 55 kilodaltons. This polypeptide was resolved to multiple isoforms by two-dimensional electrophoresis. Fractionation of protoplasts and purification of subcellular organelles has shown that enzyme activity is associated with the chloroplasts, mitochondria and cytosol (in this order, approx. 77%, 3%, and 20% of the total activity). Distinct multiple isoforms of the enzyme, which differed in isoelectric point and were compartment-specific, were resolved from purified mitochondria and chloroplasts. The latency of the glutathione reductase activity which co-purified on Percoll gradients with the mitochondrial marker enzyme, cytochrome-c oxidase (EC 1.9.3.1.), indicated that this enzyme was within the mitochondrion. The mitochondrial glutathione reductase activity was strongly dependent on NADPH and not NADH.

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Abbreviations

Da:

dalton

GR:

glutathione reductase

IEF:

isoelectric focussing

IgG:

immunoglobulin G

pI:

isoelectric point

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Authors and Affiliations

  1. John Innes Institute and AFRC Institute of Plant Science Research, Colney Lane, NR4 7UH, Norwich, UK

    E. Anne Edwards, Stephen Rawsthorne & Philip M. Mullineaux

Authors
  1. E. Anne Edwards
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  2. Stephen Rawsthorne
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  3. Philip M. Mullineaux
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Additional information

We are grateful to Dr. Alison M. Smith for advice on enzyme assays, Dr. Ian Dry for helpful suggestions during the preparation of the manuscript and Tarn Dalzell for typing the manuscript. E.A.E. acknowledges a grant from the John Innes Institute. S.E. and P.M.M. are supported by the Agricultural and Food Research Council by a grant-in-aid to the John Innes Institute.

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Edwards, E.A., Rawsthorne, S. & Mullineaux, P.M. Subcellular distribution of multiple forms of glutathione reductase in leaves of pea (Pisum sativum L.). Planta 180, 278–284 (1990). https://doi.org/10.1007/BF00194008

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