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Information on EC 1.1.1.363 - glucose-6-phosphate dehydrogenase [NAD(P)+] and Organism(s) Leuconostoc mesenteroides and UniProt Accession P11411
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1.1.1.363 glucose-6-phosphate dehydrogenase [NAD(P)+]IUBMB Comments
The enzyme catalyses a step of the pentose phosphate pathway. The enzyme from the Gram-positive bacterium Leuconostoc mesenteroides prefers NADP+ while the enzyme from the Gram-negative bacterium Gluconacetobacter xylinus prefers NAD+. cf. EC 1.1.1.49, glucose-6-phosphate dehydrogenase (NADP+) and EC 1.1.1.388, glucose-6-phosphate dehydrogenase (NAD+).
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Word Map
- 1.1.1.363
- citrate
- mesenteroides
-
leuconostoc
-
clarias
- batrachus
- 4-hydroxy-2-nonenal
-
freshwater
-
multicatalytic
- catfish
- hne
-
hne-treated
-
lipogenic
-
8-anilino-1-naphthalenesulfonic
- actinomycin
- pentose
-
disease-related
-
teleost
- synthesis
The taxonomic range for the selected organisms is: Leuconostoc mesenteroides
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
The expected taxonomic range for this enzyme is: Bacteria, Eukaryota
Synonyms
glu-6-pdh, g6-pdh, glc6pd, pputg6pdh-1, 
more
topSYNONYM 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
REACTION
REACTION DIAGRAM
COMMENTARY
ORGANISM
UNIPROT
LITERATURE
D-glucose 6-phosphate + NAD(P)+ = 6-phospho-D-glucono-1,5-lactone + NAD(P)H + H+
mechanism in which His240 acts as the general base that abstracts the proton from the C1-hydroxyl group of glucose 6-phosphate, and the carboxylate group of D177 stabilizes the positive charge that forms on H240 in the transition state. The results also confirm the postulated role of His178 in binding the phosphate moiety of glucose 6-phosphate
-
PATHWAY SOURCE
PATHWAYS
BRENDA
SYSTEMATIC NAME
IUBMB Comments
D-glucose-6-phosphate:NAD(P)+ 1-oxidoreductase
The enzyme catalyses a step of the pentose phosphate pathway. The enzyme from the Gram-positive bacterium Leuconostoc mesenteroides prefers NADP+ while the enzyme from the Gram-negative bacterium Gluconacetobacter xylinus prefers NAD+. cf. EC 1.1.1.49, glucose-6-phosphate dehydrogenase (NADP+) and EC 1.1.1.388, glucose-6-phosphate dehydrogenase (NAD+).
SUBSTRATE
PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
-
-
?
D-glucose 6-phosphate + NAD(P)+
6-phospho-D-glucono-1,5-lactone + NAD(P)H + H+
-
-
-
-
?
D-glucose 6-phosphate + NADP+ 2',3'-dialdehyde
6-phospho-D-glucono-1,5-lactone + NADPH 2',3'-dialdehyde + H+
-
-
-
-
?
D-glucose 6-phosphate + thionicotinamide-NAD+
6-phospho-D-glucono-1,5-lactone + thionicotinamide-NADH + H+
-
-
-
-
?
D-glucose 6-phosphate + thionicotinamide-NADP+
6-phospho-D-glucono-1,5-lactone + thionicotinamide-NADPH + H+
-
-
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
-
721400, 721401, 721402, 721406, 721411, 721511, 721570, 721573, 721580, 721584, 722578, 722590, 740878
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
iso ordered bi bi reaction
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
NAD+, NADP+, and pyridoxylation produce different conformational changes in the enzyme
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
steady-state random mechanism
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
the mechanism with NAD+ is more complex compared to the reaction with D-glucose 6-phosphate and NADP+
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
-
721400, 721401, 721402, 721406, 721411, 721511, 721570, 721573, 721580, 721584, 722476, 722578, 722590, 740878
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
NAD+, NADP+, and pyridoxylation produce different conformational changes in the enzyme
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
ordered bi bi reaction
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
ordered, sequential mechanism for this reaction in which NADP+ is bound first to the enzyme and NADPH released last
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
ordered, sequential mechanism. NADP+ binds first
-
-
?
NATURAL SUBSTRATE
NATURAL PRODUCT
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate)
(Substrate)
LITERATURE
(Substrate)
(Substrate)
COMMENTARY
(Product)
(Product)
LITERATURE
(Product)
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
r=reversible
ir=irreversible
?=not specified
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
-
-
?
D-glucose 6-phosphate + NADP+
6-phospho-D-glucono-1,5-lactone + NADPH + H+
-
-
-
?
D-glucose 6-phosphate + NAD+
6-phospho-D-glucono-1,5-lactone + NADH + H+
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NAD+
kcat/Km of NADP+ is 9.3fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction, kcat/Km of D-glucose 6-phosphate in the NADP+-dependent reaction is 3.5fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction. kcat/Km of D-glucose 6-phosphate in the NADP+-dependent reaction is 3.5fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction
NAD+
the NAD+/enzyme complex is half-open. It is suggested that if NAD+ approaches the open form of the enzyme, and this approach generates a conformation change, initially to the half-open form. The concomitant movement of the coenzyme sheet allows Gln47 to approach the adenine ribose 20-hydroxyl and moves away the side chain of Arg46
NADP+
binds to the furthest open form of the enzyme. Of the residues within the coenzyme domain, only Arg46 moves, interacting with the 20-phosphate and adenine. NAD+ is less well defined in the binding site
NADP+
kcat/Km of NADP+ is 9.3fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction, kcat/Km of D-glucose 6-phosphate in the NADP+-dependent reaction is 3.5fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction. kcat/Km of D-glucose 6-phosphate in the NADP+-dependent reaction is 3.5fold higher compared to kcat/Km of D-glucose 6-phosphate in NAD+-dependent reaction
NAD+
-
the maximum quenching of protein fluorescence is 50% for NAD+. The dissociation constant for NAD+ is 2.5 mM
NAD+
-
the mechanism with NAD+ is more complex compared to the reaction with D-glucose 6-phosphate and NADP+
NADP+
-
ordered, sequential mechanism for this reaction in which NADP+ is bound first to the enzyme and NADPH released last
NADP+
-
the maximum quenching of protein fluorescence is 5% for NADP+. The dissociation constant for NADP+ is 0.003 mM
NADP+
-
wild-type enzyme prefers NADP+ as coenzyme. Arg46 plays a key role in NADP+-binding
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
4-hydroxy-2-nonenal
-
pseudo first-order loss of enzyme activity. The pH dependence of the inactivation rate exhibits an inflection around pH 10, and the enzyme is protected from inactivation by glucose 6-phosphate. Loss of enzyme activity corresponds with the formation of one carbonyl function per enzyme subunit and the appearance of a lysine-4-hydroxy-2-nonenal adduct
cis-9-octadecenoyl-CoA
-
2.0 mM, 9% inhibition of NADP+-dependent reaction, 90% inhibition of NAD+-dependent reaction
citrate
-
incubation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides with Fe2+ and citrate results in rapid O2-dependent inactivation of the enzyme. The Fe(2+)-citrate complex binds to the glucose 6-phosphate binding site and then undergoes reaction with H2O2 formed in solution leading to the oxidative modification of amino acids essential for enzyme activity
CoA
-
3.4 mM, 12% inhibition of NADP+-dependent reaction, 82% inhibition of NAD+-dependent reaction
D-glucose 1-phosphate
-
a substrate-competitive inhibitor, that lowers the dissociation constant and maximum fluorescence quenching for NAD+ but not for NADP+
D-glucose 6-phosphate
-
high concentrations inhibit the NADP+-linked reaction in the dual wavelength assay (a method employing a mixture of one coenzyme and the thionicotinamide analog of the other coenzyme). Such inhibition is not observed in conventional assays using either NADP+ or thionicotinamide-NADP+
malonyl-CoA
-
2.4 mM, no inhibition of NADP+-dependent reaction, 14% inhibition of NAD+-dependent reaction
NADP+ 2',3'-dialdehyde
-
irreversible inactivation in absence of substrate. The inactivation is first order with respect to NADP+ concentration and follows saturation kinetics, indicating that the enzyme initially forms a reversible complex with the inhibitor followed by covalent modification. NADP+ and NAD+ protect the enzyme from inactivation. One molecule of NADP+ 2',3'-dialdehyde binds per subunit of glucose-6-phosphate dehydrogenase when the enzyme is completely inactivated
palmitoyl-CoA
-
inhibition is greatly diminished at high glucose 6-phosphate concentration
pyridoxal 5'-diphospho-5'-adenosine
-
inhibits competitively with respect to glucose 6-phosphate and noncompetitively with respect to NAD+ or NADP+. 0.85 mol of pyridoxal 5'-diphospho-5'-adenosine is required for complete inactivation. Lys21 and Lys343 are the sites of pyridoxal 5'-diphospho-5'-adenosine interaction. Both glucose 6-phosphate and NAD+ protect both lysyl residues against this covalent modification
vanadate
-
inhibition by vanadate dimer and tetramer. The inhibition by vanadate is competitive with respect to NAD+ or NADP+ and noncompetitive (a mixed type) with respect to glucose 6-phosphate when NAD+ or NADP+ are cofactors. The vanadate dimer is the major inhibiting species with respect to NADP+. The vanadate tetramer is the major inhibiting species with respect to glucose 6-phosphate and with respect to NAD+. No inhibition by monomeric vanadate
additional information
-
no inhibition by iodoacetate, iodoacetamide, and p-hydroxymercuribenzoate
-
acetyl-CoA
-
2.6 mM, no inhibition of NADP+-dependent reaction, 70% inhibition of NAD+-dependent reaction
acetyl-CoA
-
inhibition is greatly diminished at high glucose 6-phosphate concentration
ATP
-
3.9 mM, 50% inhibition of NADP+-dependent reaction, 90% inhibition of NAD+-dependent reaction
ATP
-
inhibition is greatly diminished at high glucose 6-phosphate concentration
ATP
-
the kinetics of ATP inhibition of the NAD+- and NADP+-linked reactions examined at pH 6.2 and pH 7.8. The results are interpreted in terms of ATP addition to binary enzyme-coenzyme and enzyme-glucose 6-phosphate complexes
Fe2+
-
incubation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides with Fe2+ and citrate results in rapid O2-dependent inactivation of the enzyme. The Fe(2+)-citrate complex binds to the glucose 6-phosphate binding site and then undergoes reaction with H2O2 formed in solution leading to the oxidative modification of amino acids essential for enzyme activity
Fe2+
-
the enzyme is rapidly inactivated by micromolar concentrations of Fe2+ and H2O2. Inactivation correlates with the formation of one carbonyl functionality/enzyme subunit, indicating that inactivation is the result of site-specific oxidative modification. Fe2+ binds to the glucose 6-phosphate binding site and interaction of the enzyme-bound Fe2+ with H2O2 leads to the oxidative modification of amino acids essential for enzyme activity. Partially inactivated enzyme remains predominantly in the dimeric form, and no change in the apparent affinity of the remaining active subunits for substrate is observed. Partial inactivation leads to a decrease in the thermal stability of the remaining activity. This decrease in thermal stability could be largely overcome by the addition of glucose 6-phosphate. Thus, although exposure to H2O2 and Fe2+ results in the irreversible inactivation of the enzyme, the resulting modification is selective, leads to the formation of heterodimers of both active and inactive subunits, and does not appear to cause large scale structural changes
H2O2
-
the enzyme is rapidly inactivated by micromolar concentrations of Fe2+ and H2O2. Inactivation correlates with the formation of one carbonyl functionality/enzyme subunit, indicating that inactivation is the result of site-specific oxidative modification. Fe2+ binds to the glucose 6-phosphate binding site and interaction of the enzyme-bound Fe2+ with H2O2 leads to the oxidative modification of amino acids essential for enzyme activity. Partially inactivated enzyme remains predominantly in the dimeric form, and no change in the apparent affinity of the remaining active subunits for substrate is observed. Partial inactivation leads to a decrease in the thermal stability of the remaining activity. This decrease in thermal stability could be largely overcome by the addition of glucose 6-phosphate. Thus, although exposure to H2O2 and Fe2+ results in the irreversible inactivation of the enzyme, the resulting modification is selective, leads to the formation of heterodimers of both active and inactive subunits, and does not appear to cause large scale structural changes
NADH
-
inhibits noncompetitively with respect to thionicotinamide-NAD+
NADH
-
product inhibition. With NAD+ as the varied substrate and glucose 6-phosphate, nonsaturating, linear noncompetitive inhibition is observed. When an experiment is performed using saturating glucose 6-phosphate, the inhibition again appears to be noncompetitive. When glucose 6-phosphate is the varied substrate inhibition by NADH appears to be noncompetitive whether NAD+ is nonsaturating or saturating
NADP+
-
increasing NADPH/NADP+ concentration ratios inhibit the NADP-linked, but stimulate the NAD-linked reaction
NADPH
-
increasing NADPH/NADP+ concentration ratios inhibit the NADP-linked, but stimulate the NAD-linked reaction
NADPH
-
inhibits competitively with respect to thionicotinamide-NADP+
NADPH
-
NADPH is tested as an alternate product inhibitor for the NAD+-linked reaction. With glucose 6-phosphate saturating and NAD+ varied the inhibition is linear noncompetitive
pyridoxal 5'-phosphate
-
inhibition in phosphate buffers at pH 7.7 and pH 6.3. 0.1 mM Pyridoxal 5-phosphate produces 50% inhibition at pH 7.7. At the lower pH more pyridoxal 5-phosphate is required to give the same degree of inhibition. The enzyme must be incubated with pyridoxal 5-phosphate prior to assaying in order to achieve maximum inhibition. Pyridoxamine 5-phosphate and pyridoxal are only slightly inhibitory. When glucose 6-phosphate is varied at a constant, nearly saturating concentration of NAD+ pyridoxal 5-phosphate behaves like a competitive inhibitor. When NAD+ is varied and glucose 6-phosphate maintained at nearly saturating concentration, inhibition by pyridoxal 5-phosphate is noncompetitive
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
NADP+
-
increasing NADPH/NADP+ concentration ratios inhibit the NADP-linked, but stimulate the NAD-linked reaction
NADPH
-
increasing NADPH/NADP+ concentration ratios inhibit the NADP-linked, but stimulate the NAD-linked reaction
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
additional information
additional information
-
the Km-value of NAD+ is insensitive to ionic strength over the range tested but that the Km for glucose 6-phosphate is affected. This effect is greatest at pH 9.0
-
0.069
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, wild-type enzyme
0.097
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21R
0.114
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, wild-type enzyme
0.239
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21R
2.5
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21Q
6.07
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21Q
0.007
D-glucose 6-phosphate
-
pH 7.8, 25°C, cosubstrate: thionicotinamide-NAD+
0.009
D-glucose 6-phosphate
-
pH 7.8, 25°C, cosubstrate: thionicotinamide-NADP+
0.063
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y415F
0.0692
D-glucose 6-phosphate
-
wild type enzyme, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.074
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y415F
0.075
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14S
0.075
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47A
0.078
D-glucose 6-phosphate
-
25°C, pH 7.8, NAD+-dependent reaction
0.0891
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.09
D-glucose 6-phosphate
-
25°C, pH 7.8, NADP+-dependent reaction
0.096
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47A
0.114
D-glucose 6-phosphate
-
wild type enzyme, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.117
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N
0.118
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14S
0.132
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme R46E
0.135
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme D374Q
0.165
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y179F
0.169
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14A
0.175
D-glucose 6-phosphate
-
mutant enzyme R46A, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.181
D-glucose 6-phosphate
-
free enzyme, with NADP+ as cosubstrate, at pH 7.5 and 37°C
0.193
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme D374Q
0.228
D-glucose 6-phosphate
-
tetraethyl orthosilicate-gel-immobilized enzyme, with NADP+ as cosubstrate, at pH 7.5 and 37°C
0.233
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47E
0.239
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47E
0.29
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N
0.334
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.378
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14A
0.441
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y179F
0.467
D-glucose 6-phosphate
-
mutant enzyme R46A, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.8
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N
0.825
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149G
1.04
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme R46E
1.08
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149V
1.28
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N
1.3
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343R
1.73
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149V
2.6
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149G
3.08
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343R
3.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182R
6.8
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182Q
13.1
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182Q
13.6
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N
16.1
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182R
25.3
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343Q
42.9
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N
106
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343Q
0.233
NAD+
-
mutant enzyme R46Q, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.00799
NADP+
-
wild type enzyme, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.854
NADP+
-
mutant enzyme R46Q, in 33 mM Tris-HCl, pH 7.6, at 25°C
5.57
NADP+
-
mutant enzyme R46A, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.011
thionicotinamide-NAD+
-
25°C, pH 7.2, cosubstrate: D-glucose 6-phosphate
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
100
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21Q
365
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21R
522
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, wild-type enzyme
582
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21R
1125
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, wild-type enzyme
1423
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21Q
0.007
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N
0.02
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N
0.6
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149V
1.1
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N
2.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149V
4.85
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N
8.5
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149G
13.8
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N
14.2
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N
17.5
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149G
75.2
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme D374Q
183
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y415F
208
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme D374Q
235
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47A
255
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182Q
263
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y179F
308
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182Q
317
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y415F
378
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14S
403
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343R
475
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y179F
522
D-glucose 6-phosphate
-
wild type enzyme, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
552
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47E
577
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47A
583
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14S
585
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182R
632
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14A
633
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme R46E
658
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme R46E
712
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343Q
750
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343R
808
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47E
808
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14A
825
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
850
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343Q
887
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
1068
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182R
1125
D-glucose 6-phosphate
-
wild type enzyme, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
1535
D-glucose 6-phosphate
-
mutant enzyme R46A, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
1800
D-glucose 6-phosphate
-
mutant enzyme R46A, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
16.5
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21Q
569
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21Q
2435
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme K21R
2956
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NADP+, wild-type enzyme
3762
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme K21R
10540
D-glucose 6-phosphate
25°C, pH 7.6, cosubstrate: NAD+, wild-type enzyme
0.005
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H240N
0.025
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H240N
0.32
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme H1787N
0.5
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149V
1.04
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme H1787N
1.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149V
3.34
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme P149G
6.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343Q
8.9
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y415F
9.4
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NADP+, mutant enzyme D177N
16.7
D-glucose 6-phosphate
-
25°C, pH 7.6, cosubstrate: NAD+, mutant enzyme D177N
21.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme P149G
23.4
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182Q
33.4
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343Q
36.7
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K182R
132
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme K343R
284
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182R
384
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme D374Q
601
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K343R
601
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Y179F
618
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme R46E
1553
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme D374Q
1670
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14A
2304
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47E
2650
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
2889
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y179F
3140
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme Q47A
3206
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NADP+, mutant enzyme T14S
3474
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47E
3814
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme K182Q
3850
D-glucose 6-phosphate
-
mutant enzyme R46A, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
4583
D-glucose 6-phosphate
-
wild type enzyme, with NADP+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
4760
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14A
4993
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme R46E
5043
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Y415F
6012
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme Q47A
7799
D-glucose 6-phosphate
-
25°C, pH 7.8, cosubstrate: NAD+, mutant enzyme T14S
8767
D-glucose 6-phosphate
-
mutant enzyme R46A, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
9267
D-glucose 6-phosphate
-
mutant enzyme R46Q, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
16250
D-glucose 6-phosphate
-
wild type enzyme, with NAD+ as cosubstrate, in 33 mM Tris-HCl, pH 7.6, at 25°C
5.57
NADP+
-
mutant enzyme R46A, in 33 mM Tris-HCl, pH 7.6, at 25°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
IMAGE
0.15
cis-9-octadecenoyl-CoA
-
24°C, pH 7.8, inhibition of NAD+-dependent reaction
0.4
malonyl-CoA
-
24°C, pH 7.8, inhibition of NAD+-dependent reaction
0.039
pyridoxal 5'-phosphate
-
25°C, pH 7.7, competitive inhibition when glucose 6-phosphate is varied at a constant, nearly saturating concentration of NAD+
additional information
vanadate
-
inhibition constants with respect to D-glucose 6-phosphate, NAD+ or NADP+
0.0062
NADP+
25°C, pH 7.6, wild-type enzyme, mutant enzyme K121R
0.0078
NADP+
25°C, pH 7.6, wild-type enzyme, mutant enzyme K21Q
0.00337
NADP+
-
wild type enzyme, in 33 mM Tris-HCl, pH 7.6, at 25°C
1.79
NADP+
-
mutant enzyme R46Q, in 33 mM Tris-HCl, pH 7.6, at 25°C
8.23
NADP+
-
mutant enzyme R46A, in 33 mM Tris-HCl, pH 7.6, at 25°C
0.034
pyridoxal 5'-diphospho-5'-adenosine
-
25°C, pH 7.8, NAD+-linked reaction
0.04
pyridoxal 5'-diphospho-5'-adenosine
-
25°C, pH 7.8, NADP+-linked reaction
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
-
a method is described which enables one to assay simultaneously the NAD+- and NADP+-linked reactions of dehydrogenases which can utilize both coenzymes. The method is based on the fact that the thionicotinamide analogs of NADH and NADPH absorb light maximally at 400 nm, a wavelength sufficiently far removed from the absorbance maximum of NADH and NADPH to permit measurements of the simultaneous reduction of NAD+ (or NADP+) and the thionicotinamide analog of NADP+ (or NAD+). Application of the method to glucosed 6-phosphate dehydrogenase from Leuconostoc mesenteroides reveals differential effects of glucose 6-phosphate concentration on the NAD+- and NADP+-linked reactions catalyzed by this enzyme which can not be detected by conventional assay procedures and which may have regulatory significance
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
additional information
log-log plots of the dependence of kcat and kcat/Km on pH for both D177N and wild-type enzyme. The kcat profile for mutant enzyme D177N shows a nearly linear increase from pH 5 until wild-type-like activity is regained at pH 10. Above pH 10 the kcat decreases precipitously. Linear regression of the data in the pH range 5-10 produces a slope of 0.9
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
pI VALUE
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
ORGANISM
COMMENTARY
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
UNIPROT
ENTRY NAME
ORGANISM
NO. OF AA
NO. OF TRANSM. HELICES
MOLECULAR WEIGHT[Da]
SOURCE
SEQUENCE
LOCALIZATION PREDICTION
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable).
The reliability class of the localization prediction ranges from 1 (very reliable) to 5 (not reliable). MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
SUBUNIT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
CRYSTALLIZATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
cocrystallization of and mutant enzyme Q365, mutant enzymen S215, mutant enzyme S215 with NAD+ and mutant enzyme Q365 with NADP+, hanging-drop vapour diffusion method, structures of NADP+- and NAD+-complexed enzymes are determined at 2.2 and 2.5 A resolution
hanging drop method, crystallization of six cysteine-containing mutants (Q56C, S61C, S132C, S215C, Q365C, S428C) of the enzyme and the successful preparation and crystallization of a heavy atom derivative of mutant enzyme S215C
hanging drop vapor diffusion technique, determination of the three-dimensional structure of the D177N mutant enzyme by X-ray cryocrystallography in the presence of NAD+ and in the presence of glucose 6-phosphate plus NADPH. The structure of a glucose 6-phosphate complex of a mutant (Q365C) with normal enzyme activity is also determined and substrate binding compared
crystallized from phosphate buffer in a form suitable for X-ray crystallographic studies. The crystals diffract to better than 2.4 A. The space group is P3(1)21 (P3(2)21), a = 105.8 A, c = 225.1 A, V = 2.18 X 10(6) A(3). The asymmetric unit probably contains a single dimer
-
the three-dimensional structure of the H240N glucose 6-phosphate dehydrogenase is determined at 2.5 A resolution. Crystals are grown in the presence of either glucose 6-phosphate and NAD+ or glucose 6-phosphate and NADP+, hanging drop vapor diffusion method with 2.27 M ammonium sulfate
-
PROTEIN VARIANTS
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
D177N
absence of a negatively charged aspartate at 177 accounts for the decrease in catalytic activity at pH 7.8
K21Q
kcat/Km for D-glucose 6-phosphate in the NADP+-dependent reaction is 76.2fold lower compared to wild-type value, kcat/Km for D-glucose 6-phosphate in the NAD+-dependent reaction is 28.5fold lower compared to wild-type value
K21R
kcat/Km for D-glucose 6-phosphate in the NADP+-dependent reaction is 1.9fold lower compared to wild-type value, kcat/Km for D-glucose 6-phosphate in the NAD+-dependent reaction is 1.4fold lower compared to wild-type value
D177N
-
kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme
D205C
-
enzyme variant with cysteine close to the dimer interface, about 30% loss of specific activity compared to wild-type enzyme shows changes in activity and the efficacy of immobilization.
D374Q
-
kcat/Km for D-glucose 6-phosphate is 7.7fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 7.3fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 12.2fold lower compared to kcat/KM of wild-type enzyme
D453C
-
enzyme variant with cysteine far from the active center, no significant loss in specific activity compared to wild-type enzyme, in contrast to wild-type enzyme, the mutant enzyme is readily immobilited
H178N
-
kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme
H250N
-
kcat/KM for D-glucose 6-phosphate (with cosubstrate NADP+) is fold lower than the value for the wild-type enzyme, kcat/KM for NADP+ is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD) is fold lower than the value for the wild-type enzyme, kcat/KM for D-glucose 6-phosphate (with cosubstrate NAD+) is fold lower than the value for the wild-type enzyme
K182Q
-
kcat/Km for D-glucose 6-phosphate is 127fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.1fold higher compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.4fold lower compared to kcat/KM of wild-type enzyme
K182R
-
kcat/Km for D-glucose 6-phosphate is 81fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.1fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.6fold lower compared to kcat/KM of wild-type enzyme
K343Q
-
kcat/Km for D-glucose 6-phosphate is 445fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 2.7fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 2.6fold lower compared to kcat/KM of wild-type enzyme
K343R
-
kcat/Km for D-glucose 6-phosphate is 23fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.2fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 2fold lower compared to kcat/KM of wild-type enzyme
L218C
-
enzyme variant with cysteine close to the active center, about 30% loss of specific activity compared to wild-type enzyme, the mutant enzyme shows almost complete immobilization but poor carrier activity
P149G
-
kcat/Km for D-glucose 6-phosphate is 890fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 120fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 448fold lower compared to kcat/KM of wild-type enzyme
P149V
-
kcat/Km for D-glucose 6-phosphate is 5933fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1265fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 3843fold lower compared to kcat/KM of wild-type enzyme
Q47A
-
kcat/Km for D-glucose 6-phosphate is 1.1fold higher compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.3fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.8fold lower compared to kcat/KM of wild-type enzyme
Q47E
-
kcat/Km for D-glucose 6-phosphate is 1.3fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 6.4fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.7fold lower compared to kcat/KM of wild-type enzyme
R46A
R46C
-
mutant glucose-6-phosphate dehydrogenases with coenzyme specificity that favors NAD+, whereas the wild-type enzyme prefers NADP+ as coenzyme
R46E
-
kcat/Km for D-glucose 6-phosphate is 4.8fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1406fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 6.7fold lower compared to kcat/KM of wild-type enzyme
R46Q
-
the enzyme's Km and Ki values for NADP+ are greatly increased (2-3 orders of magnitude)
T14A
-
kcat/Km for D-glucose 6-phosphate is 1.8fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 5.7fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 12.8fold lower compared to kcat/KM of wild-type enzyme
T14S
-
kcat/Km for D-glucose 6-phosphate is 1.1fold higher compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.6fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.5fold lower compared to kcat/KM of wild-type enzyme
Y179F
-
kcat/Km for D-glucose 6-phosphate is 4.9fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 2.2fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.7fold lower compared to kcat/KM of wild-type enzyme
Y415F
-
kcat/Km for D-glucose 6-phosphate is 1.2fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NADP+ is 1.4fold lower compared to kcat/KM of wild-type enzyme, kcat/Km for NAD+ is 1.6fold lower compared to kcat/KM of wild-type enzyme
R46A
-
mutant glucose-6-phosphate dehydrogenases with coenzyme specificity that favors NAD+, whereas the wild-type enzyme prefers NADP+ as coenzyme
R46A
-
the enzyme's Km and Ki values for NADP+ are greatly increased (2-3 orders of magnitude)
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
49
-
first order rate constant for inactivation is 0.05/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
additional information
-
the enzyme is rapidly inactivated by micromolar concentrations of Fe2+ and H2O2. Fe2+ binds to the glucose 6-phosphate binding site and interaction of the enzyme-bound Fe2+ with H2O2 leads to the oxidative modification of amino acids essential for enzyme activity. Partially inactivated enzyme remains predominantly in the dimeric form, and no change in the apparent affinity of the remaining active subunits for substrate is observed. Partial inactivation leads to a decrease in the thermal stability of the remaining activity. This decrease in thermal stability could be largely overcome by the addition of glucose 6-phosphate. Thus, although exposure to H2O2 and Fe2+ results in the irreversible inactivation of the enzyme, the resulting modification is selective, leads to the formation of heterodimers of both active and inactive subunits, and does not appear to cause large scale structural changes
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
both after non-denaturing and after denaturing electrophoretic separation (SDS-PAGE) and blotting Leuconostoc mesenteroides G6PD retains its complete catalytic activity
-
chymotrypsin inactivates. First order rate constant for inactivation is 0.02/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
-
eluted mutant enzyme D453C shows almost double the activity of the immobilized enzyme, which is consistent with 49% activity loss due to immobilization. Mutant enzyme D205C produces a 1.8-fold higher activity compared to its immobilized state. Eluted mutant enzyme L218C shows 9.9 times the activity of its immobilized state
-
pronase inactivates. First order rate constant for inactivation is 0.012/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
-
thermolysin inactivates. First order rate constant for inactivation is 0.057/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
-
trypsin inactivates. First order rate constant for inactivation is 0.025/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
-
ORGANIC SOLVENT
ORGANISM
UNIPROT
COMMENTARY
LITERATURE
guanidine-HCl
urea
-
4 M. First order rate constant for inactivation is 0.019/min. Protection by 72 mM NAD+ or by 6.3 mM glucose 6-phosphate or 72 mM NADP+
guanidine-HCl
-
between 0.9 and 1.2 M denaturant the enzyme undergoes a conformational change, exposing tryptophan residues to solvent, with some loss of secondary structure and a complete loss of enzymatic activity but without dimer dissociation to subunits. This inactive, partially unfolded, dimeric intermediate was susceptible to slow aggregation. A second equilibrium transition, reflecting extensive unfolding and dimer dissociation, occurrs at denaturant concentrations above 1.4 M. In the denaturant concentration range of 1.7-1.9 M the fluorescence change occurrs in two distinct steps. The first step involves a large, very rapid drop in fluorescence whose rate is strongly dependent on the denaturant concentration. This is followed by a small, relatively slow rise in the emission intensity, the rate of which is independent of denaturant concentration. Enzymatic activity is lost with a denaturant-concentration-dependent rate, which is approx. 3times slower than the rate of the first step in fluorescence change. A denaturation mechanism incorporating several unfolding intermediates and which accounts for all the above results is presented and discussed. While the fully unfolded enzyme regains up to 55% of its original activity upon dilution of denaturant to a concentration that would be expected to support native enzyme, denaturation intermediates are able to reactivate only minimally and in fact are found to aggregate and precipitate out of solution
guanidine-HCl
-
in 4 M guanidine hydrochloride, the dimeric enzyme dissociates to subunits and is extensively unfolded
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
4°C, 8 months, enzym variants L218C, D205C or D453C, no loss of activity
-
PURIFICATION (Commentary)
ORGANISM
UNIPROT
LITERATURE
CLONED (Commentary)
ORGANISM
UNIPROT
LITERATURE
RENATURED/Commentary
ORGANISM
UNIPROT
LITERATURE
denatured in 8 M urea and dissociated into its two inactive subunits (MW 50000 Da). Denaturation leads to an approximately 80% decrease in protein fluorescence and a 20-nm red shift in the emission maximum. Upon dilution, the urea-treated enzyme regains catalytic activity (approximately 70%). The reactivated enzyme is indistinguishable from the native enzyme based on a number of physicochemical and enzymological criteria. The kinetics of renaturation and reactivation are monitored. Reactivation is stimulated to different degrees by either the initial or delayed addition of NAD+, NADP+, or glucose 6-phosphate. During the initial, rapid phase of renaturation, approximately 3 of the enzyme's 12 histidine residues become unreactive toward diethyl pyrocarbonate; concomitant with the subsequent reactivation, approximately 7 more histidines become inaccessible to diethyl pyrocarbonate
-
in 4 M guanidine-HCl, the dimeric enzyme glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides dissociates to subunits and is extensively unfolded. Rapid dilution of this high guanidine hydrochloride concentration allowes the enzyme to partially renature. The fraction of the enzyme which does not renature aggregates and precipitates out of solution, a process which can not be substantially prevented by stabilizing additives. A renaturation mechanism is described, which involves a bi-unimolecular (subunit association-folding) reaction sequence. This mechanism involves an inactive, dimeric, glucose-6-phosphate dehydrogenase-folding intermediate
-
REF.
AUTHORS
TITLE
JOURNAL
VOL.
PAGES
YEAR
ORGANISM (UNIPROT)
PUBMED ID
SOURCE
Naylor, C.E., Gover, S.; Basak, A.K.; Cosgrove, M.S.; Levy, H.R.; Adams, M.J.
NADP+ and NAD+ binding to the dual coenzyme specific enzyme Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase: different interdomain hinge angles are seen in different binary and ternary complexes
Acta Crystallogr. Sect. D
57
635-648
2001
Leuconostoc mesenteroides (P11411)
Grove, T.H.; Ishaque, A.; Levy, H.R.
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Interaction of the enzyme with coenzymes and coenzyme analogs
Arch. Biochem. Biophys.
17
307-316
1976
Leuconostoc mesenteroides
Levy, H.R.; Daouk, G.H.; Katopes, M.A.
Regulation of coenzyme utilization by the dual nucleotide-specific glucose 6-phosphate dehydrogenase from Leuconostoc mesenteroids
Arch. Biochem. Biophys.
198
406-413
1979
Leuconostoc mesenteroides
Levy, H.R.; Christoff, M.; Ingulli, J.; Ho, E.M.
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides: revised kinetic mechanism and kinetics of ATP inhibition
Arch. Biochem. Biophys.
222
473-488
1983
Leuconostoc mesenteroides
Kurlandsky, S.B.; Hilburger, A.C.; Levy, H.R.
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides: ligand-induced conformational changes
Arch. Biochem. Biophys.
264
93-102
1988
Leuconostoc mesenteroides
Szweda, L.I.; Stadtman, E.R.
Oxidative modification of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by an iron(II)-citrate complex
Arch. Biochem. Biophys.
301
391-395
1993
Leuconostoc mesenteroides
Levy, H.R.; Vought, V.E.; Yin, X.; Adams, M.J.
Identification of an arginine residue in the dual coenzyme-specific glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides that plays a key role in binding NADP+ but not NAD+
Arch. Biochem. Biophys.
326
145-151
1996
Leuconostoc mesenteroides
Coe, E.L.; Hsu, L.H.
Acyl coenzyme A inhibition of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase: a comparison of the TPN and DPN linked reactions
Biochem. Biophys. Res. Commun.
53
66-69
1973
Leuconostoc mesenteroides
Haghighi, B.; Levy, H.R.
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Conformational transitions induced by nicotinamide adenine dinucleotide, nicotinamide adenine dinucleotide phosphate, and glucose 6-phosphate monitored by fluorescent probes
Biochemistry
21
6421-6428
1982
Leuconostoc mesenteroides
Haghighi, B.; Levy, H.R.
Glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Kinetics of reassociation and reactivation from inactive subunits
Biochemistry
21
6429-6434
1982
Leuconostoc mesenteroides
Crans, D.C., Schelble, S.M.
Vanadate dimer and tetramer both inhibit glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides
Biochemistry
29
6698-6706
1990
Leuconostoc mesenteroides
Cosgrove, M.S.; Naylor, C.; Paludan, S.; Adams, M.J.; Levy, H.R.
On the mechanism of the reaction catalyzed by glucose 6-phosphate dehydrogenase
Biochemistry
37
2759-2767
1998
Leuconostoc mesenteroides
Cosgrove, M.S.; Gover, S.; Naylor, C.E.; Vandeputte-Rutten, L.; Adams, M.J.; Levy, H.R.
An examination of the role of asp-177 in the His-Asp catalytic dyad of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase: X-ray structure and pH dependence of kinetic parameters of the D177N mutant enzyme
Biochemistry
39
15002-15011
2000
Leuconostoc mesenteroides (P11411)
Vought, V.; Ciccone, T.; Davino, M.H.; Fairbairn, L.; Lin, Y.; Cosgrove, M.S.; Adams, M.J.; Levy, H.R.
Delineation of the roles of amino acids involved in the catalytic functions of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase
Biochemistry
39
15012-15021
2000
Leuconostoc mesenteroides
Plomer, J.J.; Gafni, A.
Denaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides by guanidine hydrochloride; identification of inactive, partially unfolded, dimeric intermediates
Biochim. Biophys. Acta
1122
234-242
1992
Leuconostoc mesenteroides
Plomer, J.J.; Gafni, A.
Renaturation of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides after denaturation in 4 M guanidine hydrochloride: kinetics of aggregation and reactivation
Biochim. Biophys. Acta
1163
89-96
1993
Leuconostoc mesenteroides
Murphy, N.B.; McConnell, D.J.; Schwarz, T.F.
Expression of the gene for NAD-dependent glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides cloned in Escherichia coli K-12
J. Bacteriol.
169
334-339
1987
Leuconostoc mesenteroides
Olive, C.; Geroch, M.E.; Levy, H.R.
Glucose 6-phosphate dehydrogenase from Leuconostoc mesenteroides. Kinetic studies
J. Biol. Chem.
246
2047-2057
1971
Leuconostoc mesenteroides
Levy, H.R.; Daouk, G.H.
Simultaneous analysis of NAD- and NADP-linked activities of dual nucleotide-specific dehydrogenases. Application to Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase
J. Biol. Chem.
254
4843-4837
1997
Leuconostoc mesenteroides
Adams, M.J.; Levy, H.R.; Moffat, K.
Crystallization and preliminary x-ray data for glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides
J. Biol. Chem.
258
5867-5868
1983
Leuconostoc mesenteroides
White, B.J.; Levy, H.R.
Modification of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides with the 2',3'-dialdehyde derivative of NADP+ (oNADP+)
J. Biol. Chem.
262
1223-1229
1987
Leuconostoc mesenteroides
Lee, W.T.; Flynn, T.G.; Lyons, C.; Levy, H.R.
Cloning of the gene and amino acid sequence for glucose 6-phosphate dehydrogenase from Leuconostoc mesenteroides
J. Biol. Chem.
266
13028-13034
1991
Leuconostoc mesenteroides (P11411), Leuconostoc mesenteroides
LaDine, J.R.; Carlow, D.; Lee, W.T.; Cross, R.L.; Flynn, T.G.; Levy, H.R.
Interaction of Leuconostoc mesenteroides glucose-6-phosphate dehydrogenase with pyridoxal 5'-diphospho-5'-adenosine. Affinity labeling of Lys-21 and Lys-343
J. Biol. Chem.
266
5558-5562
1991
Leuconostoc mesenteroides
Szweda, L.I.; Stadtman, E.R.
Iron-catalyzed oxidative modification of glucose-6-phosphate dehydrogenase from Leuconostoc mesenteroides. Structural and functional changes
J. Biol. Chem.
267
3096-3100
1992
Leuconostoc mesenteroides
Szweda, L.I.; Uchida, K.; Tsai, L.; Stadtman, E.R.
Inactivation of glucose-6-phosphate dehydrogenase by 4-hydroxy-2-nonenal. Selective modification of an active-site lysine
J. Biol. Chem.
268
3342-3347
1993
Leuconostoc mesenteroides
Simons, J.R.; Mosisch, M.; Torda, A.E.; Hilterhaus, L.
Site directed immobilization of glucose-6-phosphate dehydrogenase via thiol-disulfide interchange: influence on catalytic activity of cysteines introduced at different positions
J. Biotechnol.
167
1-7
2013
Leuconostoc mesenteroides
Ravera, S.; Calzia, D.; Morelli, A.; Panfoli, I.
Oligomerization studies of Leuconostoc mesenteroides G6PD activity after SDS-PAGE and blotting
Mol. Biol.
44
472-476
2010
Leuconostoc mesenteroides
Lee, W.T.; Levy, H.R.
Lysine-21 of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase participates in substrate binding through charge-charge interaction
Protein Sci.
1
329-334
1992
Leuconostoc mesenteroides (P11411)
Adams, M.J.; Basak, A.K.; Gover, S.; Rowland, P.; Levy, H.R.
Site-directed mutagenesis to facilitate X-ray structural studies of Leuconostoc mesenteroides glucose 6-phosphate dehydrogenase
Protein Sci.
2
859-662
1993
Leuconostoc mesenteroides (P11411)
Cumana, S.; Simons, J.; Liese, A.; Hilterhaus, L.; Smirnova, I.
Immobilization of glucose 6-phosphate dehydrogenase in silica-based hydrogels: A comparative study
J. Mol. Catal. B
85-86
220-228
2013
Leuconostoc mesenteroides
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