Re‐engineering the discrimination between the oxidized coenzymes NAD+ and NADP+ in clostridial glutamate dehydrogenase and a thorough reappraisal of the coenzyme specificity of the wild‐type enzyme

@article{Capone2011ReengineeringTD,
  title={Re‐engineering the discrimination between the oxidized coenzymes NAD+ and NADP+ in clostridial glutamate dehydrogenase and a thorough reappraisal of the coenzyme specificity of the wild‐type enzyme},
  author={Marina Capone and David O. Scanlon and Joanna Griffin and Paul C. Engel},
  journal={The FEBS Journal},
  year={2011},
  volume={278}
}
Clostridial glutamate dehydrogenase mutants, designed to accommodate the 2′‐phosphate of disfavoured NADPH, showed the expected large specificity shifts with NAD(P)H. Puzzlingly, similar assays with oxidized cofactors initially revealed little improvement with NADP+, although rates with NAD+ were markedly diminished. This article reveals that the enzyme’s discrimination in favour of NAD+ and against NADP+ had been greatly underestimated and has indeed been abated by a factor of > 16 000 by the… 
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Glutamate Dehydrogenases: The Why and How of Coenzyme Specificity
  • P. Engel
  • Biology
    Neurochemical Research
  • 2013
TLDR
Structural studies show NAD+-dependent, NADP+- dependent and dual-specificity GDHs are closely related and a few site-directed mutations can reverse specificity.
Reversal of the extreme coenzyme selectivity of Clostridium symbiosum glutamate dehydrogenase
TLDR
Three mutations of glutamate dehydrogenase, previously identified as having a potential bearing on coenzyme specificity, did not engender any further shift in preference when incorporated into the triple mutant, despite having a significant effect when expressed as a single mutant.
Structure of NADP+‐dependent glutamate dehydrogenase from Escherichia coli – reflections on the basis of coenzyme specificity in the family of glutamate dehydrogenases
TLDR
These studies clarify the sequence–structure relationships in bacterial GDHs, revealing that identical residues may specify different coenzyme preferences, depending on the structural context, and primary sequence alone is therefore not a reliable guide for predicting coen enzyme specificity.
Crystal structure of NAD+-dependent Peptoniphilus asaccharolyticus glutamate dehydrogenase reveals determinants of cofactor specificity.
Protein Engineering for Nicotinamide Coenzyme Specificity in Oxidoreductases: Attempts and Challenges
TLDR
This review covers 103 enzyme engineering studies from 82 articles and evaluates the accomplishments in terms of coenzyme specificity and catalytic efficiency compared to wild type enzymes of different classes.
A General Tool for Engineering the NAD/NADP Cofactor Preference of Oxidoreductases.
TLDR
This work presents a structure-guided, semirational strategy for reversing enzymatic nicotinamide cofactor specificity that leverages the diversity and sensitivity of catalytically productive cofactor binding geometries to limit the problem to an experimentally tractable scale.
Structure-based Conversion of the Coenzyme Requirement of a Short-chain Dehydrogenase/Reductase Involved in Bacterial Alginate Metabolism*
TLDR
It is shown that the two loops determine the coenzyme requirement, and loop exchange is a promising method for conversion of coen enzymatically characterized enzymes in the SDR family.
Identification of malic enzyme mutants depending on 1,2,3-triazole moiety-containing nicotinamide adenine dinucleotide analogs.

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