Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 1.1.1.45 extracted from

  • Asada, Y.; Kuroishi, C.; Ukita, Y.; Sumii, R.; Endo, S.; Matsunaga, T.; Hara, A.; Kunishima, N.
    Dimeric crystal structure of rabbit L-gulonate 3-dehydrogenase/lambda-crystallin: insights into the catalytic mechanism (2010), J. Mol. Biol., 401, 906-920.
    View publication on PubMed

Cloned(Commentary)

Cloned (Comment) Organism
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) cells Oryctolagus cuniculus

Crystallization (Commentary)

Crystallization (Comment) Organism
purified recombinant enzyme in both apo form and NADH-bound holo form, X-ray diffraction structure determination and analysis at 1.70-1.85 A resolution Oryctolagus cuniculus

Protein Variants

Protein Variants Comment Organism
D36R site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
D36R/Q41N site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
E97Q site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Oryctolagus cuniculus
Q41N site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme Oryctolagus cuniculus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.01
-
NAD+ wild-type enzyme, pH 7.0, 25°C Oryctolagus cuniculus
0.018
-
L-gulonate with NAD+, wild-type enzyme, pH 7.0, 25°C Oryctolagus cuniculus
0.028
-
NAD+ mutant enzyme Q41N, pH 7.0, 25°C Oryctolagus cuniculus
0.054
-
NAD+ mutant enzyme E97Q, pH 7.0, 25°C Oryctolagus cuniculus
0.12
-
NADP+ mutant enzyme D36R/Q41N, pH 7.0, 25°C Oryctolagus cuniculus
0.2
-
NADP+ mutant enzyme D36R, pH 7.0, 25°C Oryctolagus cuniculus
0.67
-
NADP+ wild-type enzyme, pH 7.0, 25°C Oryctolagus cuniculus
0.86
-
NADP+ mutant enzyme Q41N, pH 7.0, 25°C Oryctolagus cuniculus
1.3
-
NAD+ mutant enzyme D36R, pH 7.0, 25°C Oryctolagus cuniculus
1.3
-
L-gulonate with NAD+, mutant enzyme Q41N, pH 7.0, 25°C Oryctolagus cuniculus
1.6
-
L-gulonate with NAD+, mutant enzyme E97Q, pH 7.0, 25°C Oryctolagus cuniculus
2.4
-
L-gulonate with NADP+, wild-type enzyme, pH 7.0, 25°C Oryctolagus cuniculus
2.6
-
NAD+ mutant enzyme D36R/Q41N, pH 7.0, 25°C Oryctolagus cuniculus
2.6
-
L-gulonate with NADP+, mutant enzyme D36R/Q41N, pH 7.0, 25°C Oryctolagus cuniculus
3.3
-
L-gulonate with NAD+, mutant enzyme D36R/Q41N, pH 7.0, 25°C Oryctolagus cuniculus
14
-
L-gulonate with NAD+, mutant enzyme D36R, pH 7.0, 25°C Oryctolagus cuniculus
14
-
L-gulonate with NADP+, mutant enzyme Q41N, pH 7.0, 25°C Oryctolagus cuniculus
18
-
L-gulonate with NADP+, mutant enzyme D36R, pH 7.0, 25°C Oryctolagus cuniculus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
36000
-
2 * 36000, recombinant enzyme, SDS-PAGE Oryctolagus cuniculus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
L-gulonate + NAD+ Oryctolagus cuniculus
-
3-dehydro-L-gulonate + NADH + H+
-
r

Organism

Organism UniProt Comment Textmining
Oryctolagus cuniculus P14755
-
-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) cells Oryctolagus cuniculus

Reaction

Reaction Comment Organism Reaction ID
L-gulonate + NAD+ = 3-dehydro-L-gulonate + NADH + H+ Cys125, Glu97, and Ser124 are putative coenzyme/substrate-binding residues, catalytic mechanism with induced-fit mechanism upon coenzyme binding and involving a network-based substrate recognition, overview Oryctolagus cuniculus

Source Tissue

Source Tissue Comment Organism Textmining
lens
-
Oryctolagus cuniculus
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
L-gulonate + NAD+
-
Oryctolagus cuniculus 3-dehydro-L-gulonate + NADH + H+
-
r
L-gulonate + NAD+ NAD+ is preferred as cofactor Oryctolagus cuniculus 3-dehydro-L-gulonate + NADH + H+
-
r
L-gulonate + NADP+ NAD+ is preferred as cofactor Oryctolagus cuniculus 3-dehydro-L-gulonate + NADPH + H+
-
r
additional information active site structure and substrate recognition, importance of the dimeric state of the enzyme, overview Oryctolagus cuniculus ?
-
?

Subunits

Subunits Comment Organism
dimer 2 * 36000, recombinant enzyme, SDS-PAGE Oryctolagus cuniculus
More the enzyme possesses two domains, the N-terminal domain with a Rossmann fold and the C-terminal domain with a helical fold. In the N-terminal domain of the NADH-bound structure exist 11 coenzyme-binding residues and two distinct side-chain conformers of coenzyme binding residue Ser124. Subunit dimerization is mediated by numerous intersubunit interactions, including 22 hydrogen bonds and 104 residue pairs of van der Waals interactions, of which those between two cognate C-terminal domains are predominant. Domain and subunit interface structures, structure comparison with the human enzyme, overview Oryctolagus cuniculus

Synonyms

Synonyms Comment Organism
More the enzyme belongs to the GDH/lambdaCRY family Oryctolagus cuniculus

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
assay at Oryctolagus cuniculus

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
7
-
assay at Oryctolagus cuniculus

Cofactor

Cofactor Comment Organism Structure
NAD+ cofactor binding mode, overview Oryctolagus cuniculus
NADH cofactor binding mode, overview Oryctolagus cuniculus
NADP+ cofactor binding mode, overview Oryctolagus cuniculus
NADPH cofactor binding mode, overview Oryctolagus cuniculus

General Information

General Information Comment Organism
physiological function L-gulonate 3-dehydrogenase is a bifunctional dimeric protein that functions not only as an NAD+-dependent enzyme in the uronate cycle but also as a taxon-specific lambda-crystallin in rabbit lens Oryctolagus cuniculus