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Conformational changes in bovine-liver glutamate dehydrogenase: a spin-label study.

A Zantema·H J Vogel·G T Robillard
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Abstract

A spin-labelled analogue of p-chloromercuribenzoate reacts specifically with glutamate dehydrogenase. The most marked change in the properties of the spin-labelled enzyme is a fivefold decrease in the rate of reduction of the coenzyme by L-glutamate and no change in the rate of oxidation by 2-oxoglutarate. The electron spin resonance spectrum is a sensitive probe for the conformational state of the enzyme. Spin-labelled glutamate dehydrogenase in the presence of saturating concentrations of NADPH and 2-oxoglutarate or L-glutamate shows a complete conformational change while in the presence of NADP+ and 2-oxoglutarate only half of the protomers have changed conformation. The conformational change upon addition of NADPH to the spin-labelled glutamate dehydrogenase in the presence of 2-oxoglutarate happens in a concerted way between 20 and 80% saturation with NADPH. One of the conformations is favoured by the activator ADP while the other is favoured by the inhibitor GTP.

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AnimalsCattleChloromercuribenzoatesGlutamate DehydrogenaseKineticsLiverMagnetic Resonance SpectroscopyNADPProtein ConformationSpin LabelsTemperatureJournal Article

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A Zantema, H J Vogel, G T Robillard. 1979-06-01. Conformational changes in bovine-liver glutamate dehydrogenase: a spin-label study.. https://doi.org/10.1111/j.1432-1033.1979.tb13058.x

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