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Literature summary for 1.1.1.12 extracted from

  • Bae, B.; Sullivan, R.P.; Zhao, H.; Nair, S.K.
    Structure and engineering of L-arabinitol 4-dehydrogenase from Neurospora crassa (2010), J. Mol. Biol., 402, 230-240.
    View publication on PubMed
Search for more literature for 1.1.1.12

Crystallization (Commentary)

Crystallization (Comment) Organism
molecular replacement method using the coordinates of human sorbitol dehydrogenase, PDB ID 1PL8. Each monomer contains a bidomain architecture composed of a large catalytic domain of residues Ala5 through Val167, and residues Arg308 through Leu362, and a smaller cofactor-binding domain with residues Ala168 through Tyr307, with a large cleft separating the two domains Neurospora crassa

Protein Variants

Protein Variants Comment Organism
D211S strong decrease in activity Neurospora crassa
D211S/I212R strong decrease in activity, increase in activity with cofactor NADP+ Neurospora crassa
D211S/I212R/D213N strong decrease in activity, increase in activity with cofactor NADP+ Neurospora crassa
D211S/I212R/S348T strong decrease in activity, increase in activity with cofactor NADP+ Neurospora crassa

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.14
-
 NAD+ wild-type, pH 8.0, 25°C Neurospora crassa
0.48
-
 NADP+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
0.55
-
 NADP+ mutant D211S/I212R/S348T, pH 8.0, 25°C Neurospora crassa
1.45
-
 NADP+ mutant D211S/I212R/D213N, pH 8.0, 25°C Neurospora crassa
2.9
-
 NAD+ mutant D211S, pH 8.0, 25°C Neurospora crassa
3.6
-
 NAD+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
5
-
 NADP+ or above, mutant D211S, pH 8.0, 25°C Neurospora crassa
5
-
 NAD+ or above, mutant D211S/I212R/D213N, pH 8.0, 25°C Neurospora crassa
5
-
 NAD+ or above, mutant D211S/I212R/S348T, pH 8.0, 25°C Neurospora crassa
8
-
 NADP+ or above, wild-type, pH 8.0, 25°C Neurospora crassa

Metals/Ions

Metals/Ions Comment Organism Structure
Zn2+ a structural zinc ion is situated at a loop region located adjacent to the catalytic domain, where it is ligated by enzyme residues Cys108, Cys111, Cys114, and Cys122. The catalytically requisite zinc ion constitutes the second metal found in each monomer of LAD. This metal is coordinated by residues Cys53, His78, and Glu79, with a water molecule completing a near-tetrahedral coordination sphere Neurospora crassa

Organism

Organism UniProt Comment Textmining
Neurospora crassa Q7SI09
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Neurospora crassa

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
adonitol + NAD+
-
 Neurospora crassa ? + NADH + H+
-
 ?
L-arabinitol + NAD+
-
 Neurospora crassa L-xylulose + NADH + H+
-
 ?
L-arabinitol + NADP+ no activity with cofactor NADP+ for wild-type Neurospora crassa L-xylulose + NADPH + H+
-
 ?
additional information no substrate: D-arabinitol. The promiscuity of LAD towards different substrates is restricted to five-carbon sugars, and no activity is observed towards either D-sorbitol or D-mannitol Neurospora crassa ?
-
 ?
xylitol + NAD+
-
 Neurospora crassa D-xylulose + NADH + H+
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.18
-
 NAD+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
8.23
-
 NADP+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
8.58
-
 NAD+ mutant D211S, pH 8.0, 25°C Neurospora crassa
10.83
-
 NAD+ wild-type, pH 8.0, 25°C Neurospora crassa
11.9
-
 NADP+ mutant D211S/I212R/D213N, pH 8.0, 25°C Neurospora crassa
20.16
-
 NADP+ mutant D211S/I212R/S348T, pH 8.0, 25°C Neurospora crassa

Cofactor

Cofactor Comment Organism Structure
NAD+ electron density corresponding to the bound NAD+ cofactor is observed within the smaller nucleotide-binding domain, along a crevice between the two domains. The adenine ring is nestled in a shallow pocket created by numerous hydrophobic residues, including Ile212, Val232, Thr260, and Val262. The nicotinamide is adjacent to the catalytic zinc ion, where it is poised for hydride transfer to the C2 atom of the substrate Neurospora crassa

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.88
-
 NAD+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
2.92
-
 NAD+ mutant D211S, pH 8.0, 25°C Neurospora crassa
8.27
-
 NADP+ mutant D211S/I212R/D213N, pH 8.0, 25°C Neurospora crassa
17.17
-
 NADP+ mutant D211S/I212R, pH 8.0, 25°C Neurospora crassa
36.6
-
 NADP+ mutant D211S/I212R/S348T, pH 8.0, 25°C Neurospora crassa
79
-
 NAD+ wild-type, pH 8.0, 25°C Neurospora crassa