Cassette mutagenesis of lysine 130 of human glutamate dehydrogenase. An essential residue in catalysis.

@article{Cho2001CassetteMO,
  title={Cassette mutagenesis of lysine 130 of human glutamate dehydrogenase. An essential residue in catalysis.},
  author={S. W. Cho and Hye-Young Yoon and J. Y. Ahn and E Y.C. Lee and J E Lee},
  journal={European journal of biochemistry},
  year={2001},
  volume={268 11},
  pages={
          3205-13
        }
}
  • S. Cho, H. Yoon, J. Lee
  • Published 1 June 2001
  • Biology, Chemistry
  • European journal of biochemistry
It has been suggested that reactive lysine residue(s) may play an important role in the catalytic activities of glutamate dehydrogenase (GDH). There are, however, conflicting views as to whether the lysine residues are involved in Schiff's base formation with catalytic intermediates, stabilization of negatively charged groups or the carbonyl group of 2-oxoglutarate during catalysis, or some other function. We have expanded on these speculations by constructing a series of cassette mutations at… 
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TLDR
Functional analyses of highly purified recombinant wild‐type hGDH2 revealed that this adaptive evolution dissociated the enzyme from GTP control, permitted regulation almost entirely by ADP and/or L‐leucine, and fine‐tuned its activity to the relatively low cellular pH that occurs in synaptic astrocytes during excitatory transmission.
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The properties of mutants in which lysine-128 has been changed to histidine (K128H) or arginine (K 128R) have substantially reduced catalytic centre activities and raised pH optima for activity, implying that lysin-128 is intimately involved in either direct or indirect interactions with all the substrates and also in catalysis.
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TLDR
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