Structural insights into domain movement and cofactor specificity of glutamate dehydrogenase from Corynebacterium glutamicum

Biochemical and Biophysical Research Communications
Hyeoncheol Francis SonKyung-Jin Kim

Abstract

Glutamate dehydrogenase (GDH) is an enzyme involved in the synthesis of amino acids by converting glutamate to α-ketoglutarate, and vice versa. To investigate the molecular mechanism of GDH, we determined a crystal structure of the Corynebacterium glutamicum-derived GDH (CgGDH) in complex with its NADP cofactor and α-ketoglutarate substrate. CgGDH functions as a hexamer, and each CgGDH monomer comprises 2 separate domains; a Rossmann fold cofactor-binding domain and a substrate-binding domain. The structural comparison between the apo- and cofactor/substrate-binding forms revealed that the CgGDH enzyme undergoes a domain movement during catalysis. In the apo-form, CgGDH exists as an open state, and upon binding of the substrate and cofactor the protein undergoes a conformation change to a closed state. Our structural study also revealed that CgGDH has cofactor specificity for NADP, but not NAD, and this was confirmed by GDH activity measurements. Residues involved in the stabilization of the NADP cofactor and the α-ketoglutarate substrate were identified, and their roles in substrate/cofactor binding were confirmed by site-directed mutagenesis experiments.

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Citations

Feb 23, 2018·Assay and Drug Development Technologies·Yanhong JinBenfang Helen Ruan
Aug 26, 2017·Bioscience, Biotechnology, and Biochemistry·Takeo Tomita
Mar 16, 2018·The Journal of Biological Chemistry·Prem PrakashPrasenjit Bhaumik
Jul 17, 2021·Biochemical and Biophysical Research Communications·Na LiMingzhu Wang
Nov 9, 2021·Proteins·Barsa Kanchan Jyotshna GodsoraPrasenjit Bhaumik

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