Functional dissection of a trigger enzyme: mutations of the bacillus subtilis glutamate dehydrogenase RocG that affect differentially its catalytic activity and regulatory properties.

Journal of Molecular Biology
Katrin GunkaJörg Stülke

Abstract

Any signal transduction requires communication between a sensory component and an effector. Some enzymes engage in signal perception and transduction, as well as in catalysis, and these proteins are known as "trigger" enzymes. In this report, we detail the trigger properties of RocG, the glutamate dehydrogenase of Bacillus subtilis. RocG not only deaminates the key metabolite glutamate to form alpha-ketoglutarate but also interacts directly with GltC, a LysR-type transcription factor that regulates glutamate biosynthesis from alpha-ketoglutarate, thus linking the two metabolic pathways. We have isolated mutants of RocG that separate the two functions. Several mutations resulted in permanent inactivation of GltC as long as a source of glutamate was present. These RocG proteins have lost their ability to catabolize glutamate due to a strongly reduced affinity for glutamate. The second class of mutants is exemplified by the replacement of aspartate residue 122 by asparagine. This mutant protein has retained enzymatic activity but has lost the ability to control the activity of GltC. Crystal structures of glutamate dehydrogenases that permit a molecular explanation of the properties of the various mutants are presented. Specificall...Continue Reading

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Citations

Dec 20, 2011·Journal of Bacteriology·Yong Heon LeeJohn D Helmann
Nov 29, 2014·Applied Microbiology and Biotechnology·Jonas E N MüllerTrygve Brautaset
Jul 22, 2020·Journal of Bacteriology·Rajesh BiswasBoris R Belitsky
Feb 6, 2017·Frontiers in Cellular and Infection Microbiology·Alejandra Carrea, Luis Diambra
Oct 28, 2019·Frontiers in Microbiology·Miriam DormeyerFabian M Commichau
Feb 7, 2020·Proceedings of the National Academy of Sciences of the United States of America·Yinggen KeShiping Wang
Jun 12, 2020·The Journal of Microbiology·Kesavan DineshkumarHuaxi Xu
Feb 6, 2020·Frontiers in Microbiology·Alejandra AlvaradoChristine Josenhans
Sep 10, 2015·Microbiology Spectrum·Fabian M Commichau, Jörg Stülke

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