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

BRENDA support

BRENDA Home
show all | hide all No of entries

Information on EC 1.1.1.45 - L-gulonate 3-dehydrogenase and Organism(s) Oryctolagus cuniculus and UniProt Accession P14755

for references in articles please use BRENDA:EC1.1.1.45
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
EC Tree
IUBMB Comments
Also oxidizes other L-3-hydroxyacids.
Specify your search results
Select one or more organisms in this record: ?
This record set is specific for:
Oryctolagus cuniculus
UNIPROT: P14755
Show additional data
Do not include text mining results
Include (text mining) results
Include results (AMENDA + additional results, but less precise)
Word Map
The taxonomic range for the selected organisms is: Oryctolagus cuniculus
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
l-gulonate 3-dehydrogenase, l-beta-hydroxyacid dehydrogenase, l-3-hydroxyacid dehydrogenase, l-gulonic acid dehydrogenase, more
SYNONYM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
L-3-aldonate dehydrogenase
-
-
-
-
L-3-aldonic dehydrogenase
-
-
-
-
L-3-hydroxyacid dehydrogenase
-
-
-
-
L-beta-hydroxy-acid-NAD-oxidoreductase
-
-
-
-
L-beta-hydroxyacid dehydrogenase
-
-
-
-
L-gulonate 3-dehydrogenase
-
-
L-gulonic acid dehydrogenase
-
-
-
-
additional information
the enzyme belongs to the GDH/lambdaCRY family
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
L-gulonate + NAD+ = 3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
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
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
oxidation
-
-
-
-
reduction
-
-
-
-
PATHWAY SOURCE
PATHWAYS
-
-
SYSTEMATIC NAME
IUBMB Comments
L-gulonate:NAD+ 3-oxidoreductase
Also oxidizes other L-3-hydroxyacids.
CAS REGISTRY NUMBER
COMMENTARY hide
9028-51-7
-
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
L-gulonate + NAD+
3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
L-gulonate + NADP+
3-dehydro-L-gulonate + NADPH + H+
show the reaction diagram
NAD+ is preferred as cofactor
-
-
r
? + NADP+
? + NADPH
show the reaction diagram
-
-
-
-
?
acetoacetate + NADH
? + NAD+
show the reaction diagram
-
-
-
-
?
L-3-hydroxybutyrate + ?
?
show the reaction diagram
-
-
-
-
?
L-gulonate + NAD+
3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
-
-
-
-
?
L-threonate + ?
?
show the reaction diagram
-
-
-
-
?
additional information
?
-
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
L-gulonate + NAD+
3-dehydro-L-gulonate + NADH + H+
show the reaction diagram
-
-
-
r
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
cofactor binding mode, overview
NADH
cofactor binding mode, overview
NADP+
cofactor binding mode, overview
NADPH
cofactor binding mode, overview
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Cibacron blue
-
competitive inhibition with respect to NAD+
malonate
-
reversible inhibition, IC50: 0.34 mM
malonic acid monoethyl ester
-
weak inhibition, IC50: 5.5 mM
p-chloromercuriphenylsulfonate
-
0.001 mM, complete inhibition
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.018 - 18
L-gulonate
0.01 - 2.6
NAD+
0.12 - 0.86
NADP+
8.3 - 9.9
acetoacetate
2.2 - 2.3
L-3-hydroxybutyrate
0.18 - 18.5
L-gulonate
1.2 - 2.9
L-threonate
0.01 - 0.011
NAD+
0.0005 - 0.0006
NADH
0.67 - 0.89
NADP+
additional information
additional information
-
no activity with 5-40 mM sugars or hydroxyacids like 2-keto-L-guluonate, 3-deoxyglucosone, xylitol, L-threitol, ascorbic acid, malic acid, L-threonine, L-glycerate, L-tartarate and L-lactate, or 50 mM ethyl and methyl esters of 3-hydroxybutyrate
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.34
malonate
Oryctolagus cuniculus
-
reversible inhibition, IC50: 0.34 mM
5.5
malonic acid monoethyl ester
Oryctolagus cuniculus
-
weak inhibition, IC50: 5.5 mM
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.011
-
in the supernatant
6.71
-
after purification
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
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
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION?
CRYL1_RABIT
319
0
35202
Swiss-Prot
other Location (Reliability: 3)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
36000
2 * 36000, recombinant enzyme, SDS-PAGE
35000
-
2 * 35000
36000
70000
-
gel filtration
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
dimer
2 * 36000, recombinant enzyme, SDS-PAGE
dimer
-
2 * 35000
monomer
-
X-ray diffraction, natural active protein is presumably a dimer 2 * 36000
additional information
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
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
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D36R
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D36R/Q41N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
E97Q
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
Q41N
site-directed mutagenesis, the mutant shows reduced activity compared to the wild-type enzyme
D36R
-
mutation leads to a switch in the favor of NADP(H) specificity, suggesting an important role of Asp36 in the coenzyme specificity
E157Q
-
mutation produces no large kinetic alterations
H145Q
-
inactive enzyme form
N195D
-
inactive enzyme form
N196Q
-
inactive enzyme form
S124A
-
mutation decreases the catalytic efficiency 500 fold
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
recombinant wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) cells
ammonium sulfate fractionation, Sephadex G-100 column chromatography, Q-Sepharose column chromatography, Blue-Sepharose column chromatography, hydroxylapatite column chromatography
-
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) cells
expressed in Escherichia coli
-
in Escherichia coli strain BL21(DE3)
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Ishikura, S.; Usami, N.; Araki, M.; Hara, A.
Structural and functional characterization of rabbit and human L-gulonate 3-dehydrogenase
J. Biochem.
137
303-314
2005
Oryctolagus cuniculus
Manually annotated by BRENDA team
Asada, Y.; Kuroishi, C.; Ukita, Y.; Sumii, R.; Endo, S.; Matsunaga, T.; Hara, A.; Kunishima, N.
Crystallization and preliminary X-ray crystallographic analysis of rabbit L-gulonate 3-dehydrogenase
Acta Crystallogr. Sect. F
64
228-230
2008
Oryctolagus cuniculus
Manually annotated by BRENDA team
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
J. Mol. Biol.
401
906-920
2010
Oryctolagus cuniculus (P14755), Oryctolagus cuniculus
Manually annotated by BRENDA team