Literature summary for 1.2.1.61 extracted from

Su, J.; Zhang, C.; Zhang, J.J.; Wei, T.; Zhu, D.; Zhou, N.-Y.; Gu, Lc.
Crystal structure of the gamma-hydroxymuconic semialdehyde dehydrogenase from Pseudomonas sp. strain WBC-3, a key enzyme involved in para-Nitrophenol degradation (2013), BMC Struct. Biol., 13, 30 .

Cloned(Commentary)

Cloned (Comment)	Organism
gene pnpE, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)	Pseudomonas sp. WBC-3

Crystallization (Commentary)

Crystallization (Comment)	Organism
purified recombinant enzyme PnpE in apo-form or in complex with NAD+, sitting drop vapor diffusion technique, 10 mg/ml apo-enzyme from 20% w/v PEG 3350, and 0.2 M KNO3 at 20°C, 10 mg/ml NAD+-complexed enzyme from 0.1 M bicine, pH 8.5, 20% PEG 10000, and 1 mM NAD+, X-ray diffraction structure determination and analysis at 2.7-3.1 A resolution, molecular replacement method using bovine mitochondrial aldehyde dehydrogenase, PDB ID 1A4Z, as the search model	Pseudomonas sp. WBC-3

Crystallization (Comment)

Organism

purified recombinant enzyme PnpE in apo-form or in complex with NAD+, sitting drop vapor diffusion technique, 10 mg/ml apo-enzyme from 20% w/v PEG 3350, and 0.2 M KNO3 at 20°C, 10 mg/ml NAD+-complexed enzyme from 0.1 M bicine, pH 8.5, 20% PEG 10000, and 1 mM NAD+, X-ray diffraction structure determination and analysis at 2.7-3.1 A resolution, molecular replacement method using bovine mitochondrial aldehyde dehydrogenase, PDB ID 1A4Z, as the search model

Pseudomonas sp. WBC-3

Protein Variants

Protein Variants	Comment	Organism
C281A	site-directed mutagenesis	Pseudomonas sp. WBC-3
E247K	site-directed mutagenesis	Pseudomonas sp. WBC-3
F150A	site-directed mutagenesis	Pseudomonas sp. WBC-3
F154A	site-directed mutagenesis	Pseudomonas sp. WBC-3
F447A	site-directed mutagenesis	Pseudomonas sp. WBC-3
H275E	site-directed mutagenesis	Pseudomonas sp. WBC-3
I282D	site-directed mutagenesis	Pseudomonas sp. WBC-3
N149A	site-directed mutagenesis	Pseudomonas sp. WBC-3
W157A	site-directed mutagenesis	Pseudomonas sp. WBC-3

Natural Substrates/ Products (Substrates)

Natural Substrates	Organism	Comment (Nat. Sub.)	Natural Products	Comment (Nat. Pro.)	Rev.	Reac.
4-hydroxymuconic semialdehyde + NAD+ + H2O	Pseudomonas sp. WBC-3	-	maleylacetate + NADH + 2 H+	-	?

Organism

Organism	UniProt	Comment	Textmining
Pseudomonas sp. WBC-3	C1I208	-	-

Purification (Commentary)

Purification (Comment)	Organism
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity and anion exchange chromatography, followed by gel filtration	Pseudomonas sp. WBC-3

Reaction

Reaction	Comment	Organism	Reaction ID
4-hydroxymuconic semialdehyde + NAD+ + H2O = maleylacetate + NADH + H+	catalytic mechanism involving residues Glu247 and Cys281	Pseudomonas sp. WBC-3	a

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.	Reac.
4-hydroxymuconic semialdehyde + NAD+ + H2O	-	Pseudomonas sp. WBC-3	maleylacetate + NADH + 2 H+	-	?

Subunits

Subunits	Comment	Organism
More	the monomer of PnpE has the typical structural organization betaalphabeta of the ALDH family, with 17 beta-strands and 14 alpha-helices, the secondary structures belong to three domains: a substrate binding domain, a cofactor binding domain, and an oligomerization domain mediating protein dimerization. The substrate binding domain consists of a central six-stranded beta-sheet and six alpha-helices, while the cofactor binding domain contains a nine-stranded beta-sheet and seven alpha-helices, structure, overview	Pseudomonas sp. WBC-3

Synonyms

Synonyms	Comment	Organism
gamma-hydroxymuconic semialdehyde dehydrogenase	-	Pseudomonas sp. WBC-3
NAD dependent gamma-hydroxymuconic semialdehyde dehydrogenase	-	Pseudomonas sp. WBC-3
pnpE	-	Pseudomonas sp. WBC-3

Temperature Optimum [°C]

Temperature Optimum [°C]	Temperature Optimum Maximum [°C]	Comment	Organism
25	-	assay at	Pseudomonas sp. WBC-3

pH Optimum

pH Optimum Minimum	pH Optimum Maximum	Comment	Organism
8	-	assay at	Pseudomonas sp. WBC-3

Cofactor

Cofactor	Comment	Organism	Structure
NAD+	NAD+ is situated in a cleft of PnpE. The cofactor binding site is composed of two pockets. The adenosine and the first ribose group of NAD bind in one pocket and the nicotinamide ring in the other. Six amino acids, C281, E247, Q210, W148, I146 and K172, interact with the cofactor	Pseudomonas sp. WBC-3

General Information

General Information	Comment	Organism
additional information	highly conserved residues C281 and E247 are identified to be critical for its catalytic activity, and flexible docking studies of the enzyme-substrate system predict the interactions between PnpE and its substrate 4-hydroxymuconic semialdehyde, flexible docking of substrate to PnpE, molecular docking analysis, overview	Pseudomonas sp. WBC-3
physiological function	the pnpE-encoded enzyme gamma-hydroxymuconic semialdehyde dehydrogenase catalyzes the reduction of 4-hydroxymuconic semialdehyde to maleylacetate in Pseudomonas sp. strain WBC-3, playing a key role in the catabolism of toxic para-nitrophenol to Krebs cycle intermediates	Pseudomonas sp. WBC-3