The Active Site Architecture of a Short-Chain Dehydrogenase Defined by Site-Directed Mutagenesis and Structure Modeling

Biochemistry ◽  
1994 ◽  
Vol 33 (23) ◽  
pp. 7047-7055 ◽  
Author(s):  
Lluis Ribas de Pouplana ◽  
Linda A. Fothergill-Gilmore
Keyword(s):  
Active Site ◽  
Site Directed Mutagenesis ◽  
Short Chain ◽  
Directed Mutagenesis ◽  
Structure Modeling ◽  
Short Chain Dehydrogenase

Crystal structures and functional studies clarify substrate selectivity and catalytic residues for the unique orphan enzyme N-acetyl-D-mannosamine dehydrogenase

2014 ◽  
Vol 462 (3) ◽  
pp. 499-511 ◽  
Author(s):  
Agustín Sola-Carvajal ◽  
Fernando Gil-Ortiz ◽  
Francisco García-Carmona ◽  
Vicente Rubio ◽  
Álvaro Sánchez-Ferrer
Keyword(s):  
Substrate Specificity ◽  
Crystal Structures ◽  
Functional Site ◽  
Site Directed Mutagenesis ◽  
Short Chain ◽  
Directed Mutagenesis ◽  
Substrate Selectivity ◽  
Functional Studies ◽  
Short Chain Dehydrogenase ◽  
Orphan Enzyme

Functional site-directed mutagenesis and crystallographic studies with N-acetyl-D-mannosamine dehydrogenase reveal a homotetramer, a short-chain dehydrogenase/reductase subunit fold and a highly developed C-terminal tail interlinking subunit. The structures of the complexes explain substrate specificity. The four residues forming the catalytic tetrad are identified.

scholarly journals Threonine 57 is required for the post-translational activation ofEscherichia coliaspartate α-decarboxylase

2014 ◽  
Vol 70 (4) ◽  
pp. 1166-1172 ◽  
Author(s):  
Michael E. Webb ◽  
Briony A. Yorke ◽  
Tom Kershaw ◽  
Sarah Lovelock ◽  
Carina M. C. Lobley ◽  
...  
Keyword(s):  
Crystal Structure ◽  
Active Site ◽  
Site Directed Mutagenesis ◽  
Intramolecular Rearrangement ◽  
Directed Mutagenesis ◽  
Vitamin B ◽  
Biosynthesis Pathway ◽  
Serine Residue ◽  
Translational Activation

Aspartate α-decarboxylase is a pyruvoyl-dependent decarboxylase required for the production of β-alanine in the bacterial pantothenate (vitamin B5) biosynthesis pathway. The pyruvoyl group is formedviathe intramolecular rearrangement of a serine residue to generate a backbone ester intermediate which is cleaved to generate an N-terminal pyruvoyl group. Site-directed mutagenesis of residues adjacent to the active site, including Tyr22, Thr57 and Tyr58, reveals that only mutation of Thr57 leads to changes in the degree of post-translational activation. The crystal structure of the site-directed mutant T57V is consistent with a non-rearranged backbone, supporting the hypothesis that Thr57 is required for the formation of the ester intermediate in activation.

Enzyme activity enhancement of chondroitinase ABC I from Proteus vulgaris by site-directed mutagenesis

RSC Advances ◽  
2015 ◽  
Vol 5 (93) ◽  
pp. 76040-76047 ◽  
Author(s):  
Zhenya Chen ◽  
Ye Li ◽  
Yue Feng ◽  
Liang Chen ◽  
Qipeng Yuan
Keyword(s):  
Active Site ◽  
Simulation Analysis ◽  
Docking Simulation ◽  
Site Directed Mutagenesis ◽  
Directed Mutagenesis ◽  
Wild Type ◽  
Proteus Vulgaris ◽  
Molecular Docking Simulation ◽  
Activity Enhancement ◽  
First Time

Arg660 was found as a new active site and Asn795Ala and Trp818Ala mutants showed higher activities than the wild type based on molecular docking simulation analysis for the first time.

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