Title : Importance of glutamate 279 for the coenzyme binding of human glutamate dehydrogenase.

Pub. Date : 2002 Nov 1

PMID : 12193607






5 Functional Relationships(s)
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1 Photoaffinity labeling with a specific probe, [(32)P]nicotinamide 2-azidoadenosine dinucleotide, was used to identify the NAD(+) binding site within human GDH encoded by the synthetic human GDH gene and expressed in Escherichia coli as a soluble protein. NAD glutamate dehydrogenase 1 Homo sapiens
2 Photoaffinity labeling with a specific probe, [(32)P]nicotinamide 2-azidoadenosine dinucleotide, was used to identify the NAD(+) binding site within human GDH encoded by the synthetic human GDH gene and expressed in Escherichia coli as a soluble protein. NAD glutamate dehydrogenase 1 Homo sapiens
3 Amino acid sequencing and compositional analysis identified Glu(279) as the site of photoinsertion into human GDH, suggesting that Glu(279) is located at or near the NAD(+) binding site. NAD glutamate dehydrogenase 1 Homo sapiens
4 The K(m) values for NAD(+) were 10-14-fold greater for the mutant GDHs than for wild-type GDH, whereas the V(max) values were similar for wild-type and mutant GDHs. NAD glutamate dehydrogenase 1 Homo sapiens
5 The results with photoaffinity labeling and cassette mutagenesis studies suggest that Glu(279) plays an important role for efficient binding of NAD(+) to human GDH. NAD glutamate dehydrogenase 1 Homo sapiens