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Glutamate dehydrogenase from Pisum sativum L.

Localization of the multiple forms and of glutamate formation in isolated mitochondria

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Abstract

A 2–8-fold increase in the activity of glutamate dehydrogenase (GDH), accompanied by an alteration of the GDH isoenzyme pattern, was observed in detached pea shoots floated on tap water (preincubated shoots). Sugars supressed the process, whereas NH 4+ and various metabolites as well as inhibitors of energy metabolism and protein synthesis were ineffective. The subcellular distribution pattern revealed evidence that the GDH isoenzymes are exclusively located in the mitochondrial matrix. The alterations in GDH activity occurring in preincubated shoots are restricted to the mitochondria.

An experimental device suitable for studying the GDH function in isolated intact mitochondria has been established. Using [14C] citrate as the carbon source and hydrogen donor, the mitochondria synthesized considerable amounts of glutamate upon addition of NH 4+ . The rates of glutamate formation in dependency of increasing NH 4+ levels follow simple Michaelis-Menten kinetics. Half-saturation concentrations of NH 4+ of 3.6±1.2 mM; 1.9±0.06 mM and 1.6±0.1 mM were calculated for the mitochondria isolated from pea shoots, roots, and preincubated shoots, respectively. The results are discussed in relation to the possible role of GDH in NH+/4 assimilation at elevated intracellular NH+/4 levels.

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Abbreviations

GDH:

Glutamate dehydrogenase

MDH:

malate dehydrogenase

GOT:

aspartate aminotransferase

SDH:

succinate dehydrogenase

HEPES:

4-(2-hydroxyethyl)-1-piperazineethan-sulfonic acid

BSA:

bovine serum albumin

TPP:

thiamine pyrophosphate

DNP:

2,4-dinitrophenol

CCCP:

carbonyl cyanide m-chlorophenylhydrazone

DCPIP:

2,6-dichlorophenolindophenol

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Authors and Affiliations

  1. Institut für Pharmazeutische Biologie der Technischen Universität Braunschweig, D-3300, Braunschweig, Federal Republic of Germany

    Winfried Nauen & Thomas Hartmann

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  1. Winfried Nauen
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  2. Thomas Hartmann
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Dedicated to Professor Dr. Maximilian Steiner on the occasion of his 75th birthday

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Nauen, W., Hartmann, T. Glutamate dehydrogenase from Pisum sativum L.. Planta 148, 7–16 (1980). https://doi.org/10.1007/BF00385435

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  • DOI: https://doi.org/10.1007/BF00385435

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