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C155S
nuclear relocalization of GAPC1 under cadmium-induced oxidative stress is stimulated, rather than inhibited, by mutation of the catalytic cysteine C155
C159S
the mutant C159S of the isozyme GapC2 shows decreased specific activity
D32A
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activity of mutant enzyme D32A with NAD+ is equal to that of the wild-type enzyme, mutant enzyme also shows activity with NADP+, about 3% of the activity with NAD+
D32A/L187N
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wild-type enzyme has no activity with NADP+, the mutant enzyme D32A/L187N shows catalytic efficiency with NADP+ higher than that with NAD+
L187N
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activity of mutant L187N with NAD+ is higher than that of the wild-type enzyme, mutant enzyme also shows activity with NADP+, about 7% of the activity with NAD+
D35G
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mutation enables GAPDH to accept both NAD and NADP
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D35G/L36R/P192S
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mutant accepts both Nad AND nadp WITH SIMILAR EFFICINCY
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D35G/L36T/T37K
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catalytic efficiency with NADP is about 10fold hihger than with NAD
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D35G/L36T/T37K/P192S
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Mutant shows the highest catalytic efficiency with NADP while the catalytic efficiency with NAD also increases
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L36T
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mutation enables GAPDH to accept both NAD and NADP
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C153A
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site-directed mutagenesis
N313T/Y317G
dissociation constant for NAD+ is 300times higher than that of the wild-type enzyme. Conformational equilibrium between the syn and the anti forms with a preference for the anti conformer
Y317A
dissociation constant for NAD+ is 5times higher than that of the wild-type enzyme. Wild-type syn orientation of bound nicotinamide remains unchanged
Y317G
dissociation constant for NAD+ is 13times higher than that of the wild-type enzyme. Wild-type syn orientation of bound nicotinamide remains unchanged
C149A
mutant enzyme displays no significant dehydrogenase activity
C149S
low but significant phosphorylating dehydrogenase activity
C149selenocysteine
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mutant enzyme has selenoperoxidase activity
D186G
behavior in NAD+ binding is similar to that of the wild type enzyme
D186G/E276G
positive cooperativity in binding the coenzyme NAD+
D282G
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enzyme exists as dimer and tetramer, the tetramer is inactive, the dimer is slightly active, 650fold decrease in turnover number for NAD+, 5.8fold increase in Km-value for NAD+ compared to wild-type enzyme
E276G
behavior in NAD+ binding is similar to that of the wild type enzyme
L187A/P188S
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the mutant is catalytically active not only with NAD+, as the wild-type enzyme, but also with NADP+
N313T
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mutant enzyme with a drastic decrease in thermostability, weakening of cooperative interactions between the catalytic and the cofactor domains and an inefficient binding of NAD+, mutant enzyme exists only as tetramer, 65fold decrease in turnover number for NAD+, 50fold increase in Km-value for NAD+ compared to wild-type enzyme
T34Q/T39S/L43Q
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drastic decrease in thermostability, inefficient NAD+ binding, enzyme exists as dimer and tetramer, the tetramer is inactive, the dimer is slightly active, 650fold decrease in turnover number for NAD+, 4fold increase in Km-value for NAD+ compared to wild-type enzyme
W310F
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mutant enzyme with a drastic decrease in thermostability, mutant enzyme exists only as tetramer, 2fold increase of Km-value for NAD+, 1.3fold decrease in turnover number compared to wild-type enzyme
W84F
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slightly lower Km-values for NAD+ and glyceraldehyde 3-phosphate, slightly higher Km-value for phosphate. The construction of the mutant permitts the identification of the individual fluorescence and phosphorescence characteristics of the two Trp residues W84 and W310 in the native enzyme
Y283V
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mutant enzyme with a drastic decrease in thermostability, dimeric form is inactive, KM-value and turnover-number of tetramer are nearly identical to that of the wild-type
Y46G
behavior in NAD+ binding is similar to that of the wild type enzyme
Y46G/R52G
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inactive mutant enzyme, only exists as dimer
C149A
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mutant has almost completely lost the ability to bind telomere. Upon expression in A-549 cells, mutant localizes to the nucleus but is unable to confer any significant protection of telomeres against chemotherapy-induced degradation or growth inhibition
C152G
mutant retains the ability to interact with but is unable to reactivate DNA repair enzyme APE1
C156G
mutant retains the ability to interact with but is unable to reactivate DNA repair enzyme APE1
D234A
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site-directed mutagenesis
D256R/K260E
site-directed mutagenesis, the double mutation of GAPDH results in loss of detectable binding activity to wild-type capsid N-terminal domain
D256R/K260E/K263E/E267R
site-directed mutagenesis, multiple-substituted GAPDH mutant D256R/K260E/K263E/E267R retains the oligomeric formation with wild-type GAPDH in HIV-1 producing cells, but the incorporation level of the hetero-oligomer is decreased in viral particles. The viruses produced from cells expressing the D256R/K260E/K263E/E267R mutant restores tRNALys3 packaging efficiency because the mutant exerts a dominant negative effect by preventing wild-type GAPDH from binding to matrix region and capsid N-terminal domain and improves the reverse transcription
D256R/K260E/Q264A
site-directed mutagenesis, the mutant lacks the ability to bind to the wild-type capsid N-terminal domain
D256R/K260E/Q264A/E267R
site-directed mutagenesis, the mutant lacks the binding ability to the wild-type capsid N-terminal domain
D32A
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mutant is unable to bind NAD+, is enzymatically inactive and has almost completely lost the ability to bind telomere. Upon expression in A-549 cells, mutant localizes to the nucleus but is unable to confer any significant protection of telomeres against chemotherapy-induced degradation or growth inhibition
D356R
site-directed mutagenesis, the mutation leads to loss of the ability to bind to wild-type matrix region
E267R
site-directed mutagenesis, the mutation leads to loss of the ability to bind to wild-type matrix region
H179A
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site-directed mutagenesis, the KD value of cADPR to GAPDHHis179Ala mutant protein is markedly increased compared to wild-type GAPDH enzyme
K263E
site-directed mutagenesis, the mutation leads to loss of the ability to bind to wild-type matrix region
P111A
site-directed mutagenesis, mutation at first position of alpha-helix
P157A
site-directed mutagenesis, mutation at first position of alpha-helix
P164A
site-directed mutagenesis, mutation at beta-turn, the mutant shows reduced thermostability and reduced resistance against guanidine hydrochloride. The Tm value of the heat-absorption curve decreases by 3.3°C compared to the wild-type protein
P197A
site-directed mutagenesis, mutation at beta-turn
P213A
site-directed mutagenesis, mutation at beta-turn
P326A
site-directed mutagenesis, mutation at first position of alpha-helix, the mutant shows reduced thermostability and reduced resistance against guanidine hydrochloride. The Tm value of the heat-absorption curve decreases by 6.0°C compared to the wild-type protein
A229V
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
C281W
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
G166D
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
I308S
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
V239I
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
Y327N
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exhibits 50-55% residual activity in blood compared to the wild type enzyme
C158A
site-directed mutagenesis, inactive mutant
C162A
site-directed mutagenesis, the mutant exhibits a comparable Vmax to the wild-type enzyme and only a 2fold increased Km value for D-glyceraldehyde 3-phosphate
H185A
site-directed mutagenesis, inactive mutant
N142S
naturally occuring mutation, resulting in a non-significant structural change, since Ser at position 142 also shows a similar characteristic to the wild-type residues N142, experimentally the mutation results in a loss of enzyme activity
P295L
naturally occuring mutation
N142S
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naturally occuring mutation, resulting in a non-significant structural change, since Ser at position 142 also shows a similar characteristic to the wild-type residues N142, experimentally the mutation results in a loss of enzyme activity
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P295L
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naturally occuring mutation
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C158A
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site-directed mutagenesis, inactive mutant
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C162A
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site-directed mutagenesis, the mutant exhibits a comparable Vmax to the wild-type enzyme and only a 2fold increased Km value for D-glyceraldehyde 3-phosphate
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H185A
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site-directed mutagenesis, inactive mutant
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N142S
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naturally occuring mutation, resulting in a non-significant structural change, since Ser at position 142 also shows a similar characteristic to the wild-type residues N142, experimentally the mutation results in a loss of enzyme activity
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P295L
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naturally occuring mutation
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C149S
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treatment of with (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide at 0.1 mM leads to low levels of aggregation (5% of wild type)
C149S/C281S
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mutant shows a complete absence of aggregation in the presence of (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide
C153S
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aggregation can be detected at low concentrations of (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (0.001 mM) and is enhanced at higher concentrations
C244A
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aggregation can be detected at low concentrations of (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide (0.001 mM) and is enhanced at higher concentrations
C281S
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the levels of aggregation in C281S are reduced to 45% of wild type at 0.1 mM (E)-4-ethyl-2-[(E)-hydroxyimino]-5-nitro-3-hexenamide
F37G
the mutant shows strongly reduced kcat values compared to the wild type enzyme
F37L
the mutant shows strongly reduced kcat values compared to the wild type enzyme
F37T
the mutant shows strongly reduced kcat values compared to the wild type enzyme
F37G
substitution of residue F37 with Gly, Thr or Leu leads to 6- to 9fold increase in Km value for cofactor NAD+ or NADG, with only slight increase for substrates D- glyceraldehyde 3-phosphate or 3-phospho-D-glyceroyl phosphate
F37L
substitution of residue F37 with Gly, Thr or Leu leads to 6- to 9fold increase in Km value for cofactor NAD+ or NADG, with only slight increase for substrates D-glyceraldehyde 3-phosphate or 3-phospho-D-glyceroyl phosphate
F37T
substitution of residue F37 with Gly, Thr or Leu leads to 6- to 9fold increase in Km value for cofactor NAD+ or NADG, with only slight increase for substrates D-glyceraldehyde 3-phosphate or 3-phospho-D-glyceroyl phosphate
T227A
mutation at site of O-GlcNAcylation. Mutation induces the cytoplasmic accumulation of glyceraldehyde 3-phosphate dehydrogenase
C151/H178N
the rate of the forward reaction is decreased by 47000 times compared to the wild type enzyme with similar affinity for the substrate and the coenzyme
C151G
the mutant is completely inactive
C151S
the mutant shows drastically reduced kcat values compared to the wild type enzyme
H178N
the mutant shows no significant difference in the Km values of D-glyceraldehyde 3-phosphate, phosphate, and NAD+ compared to the wild type enzyme, however, the mutation results in the reduction of kcat of oxidative phosphorylation by 1400times
C151/H178N
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the rate of the forward reaction is decreased by 47000 times compared to the wild type enzyme with similar affinity for the substrate and the coenzyme
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C151G
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the mutant is completely inactive
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C151S
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the mutant shows drastically reduced kcat values compared to the wild type enzyme
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H178N
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the mutant shows no significant difference in the Km values of D-glyceraldehyde 3-phosphate, phosphate, and NAD+ compared to the wild type enzyme, however, the mutation results in the reduction of kcat of oxidative phosphorylation by 1400times
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C152S
site-directed mutagenesis, mutation of the catalytic residue inactivates the enzyme
S124A
site-directed mutagenesis, the mutant is phosphorylated in a similar way like the wild-type enzyme, but shows highly reduced activity
S205A
site-directed mutagenesis, the mutant is poorly or not phosphorylated, the mutant shows similar affinity for both substrates but near half of the Vmax compared to wild-type
S205D
site-directed mutagenesis, the mutant enzyme (mimicking the phosphorylated form) exhibits a sstrong decrease in activity but similar affinity toward substrates compared to wild-type. The catalytic efficiency is 330 and 410fold lower with NAD+ and Ga3P, respectively
S66A
site-directed mutagenesis, the mutant is phosphorylated in a similar way like the wild-type enzyme, the mutant shows similar affinity for both substrates but near half of the Vmax compared to wild-type
C149S
the mutant shows 92% activity compared to the wild type enzyme
C149S
about 1% of wild-type activity, residue Cys149 is both essential for catalysis and the only accessible cysteine
C153S
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mutant, having only one cysteine in the catalytic site, is oxidized and glutathionylated similarly to the wild-type enzyme
C153S
the mutant shows 0.7% activity compared to the wild type enzyme
C153S
residue Cys153 has apparently no role in catalysis, in spite of the proximity in space with catalytic Cys149
D35G
mutation enables GAPDH to accept both NAD and NADP
D35G
site-directed mutagenesis, the mutant enzyme accepts both NAD+ and NADP+, the catalytic efficiency with NADP+ is 3fold lower than with NAD+
D35G/L36R/P192S
mutant accepts both Nad AND nadp WITH SIMILAR EFFICINCY
D35G/L36R/P192S
site-directed mutagenesis, the mutant enzyme accepts both NAD+ and NADP+ with similar catalytic efficiency
D35G/L36T/T37K
catalytic efficiency with NADP is about 10fold hihger than with NAD
D35G/L36T/T37K
site-directed mutagenesis, introducing a third mutation T37K into the mutant D35G/L36T completely reverses the coenzyme specificity of the enzyme
D35G/L36T/T37K/P192S
Mutant shows the highest catalytic efficiency with NADP while the catalytic efficiency with NAD also increases
D35G/L36T/T37K/P192S
site-directed mutagenesis, the mutant shows high catalytic efficiency with NADP+ while the catalytic efficiency with NAD+ also increases. The replacement of Pro192 to Ser benefits the binding affinity of both NAD+ and NADP+
L36T
mutation enables GAPDH to accept both NAD and NADP
L36T
site-directed mutagenesis, the mutant enzyme accepts both NAD+ and NADP+, the catalytic efficiency with NADP+ is lower than with NAD+
C153S
about 450fold decrease in activity
C153S
the specific activity of the mutant enzyme is approximately 450fold lower than that of the wild type enzyme
C149A
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site-directed mutagenesis
C149A
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mutation abolishes NAD+-dependent ADP-ribosylation of the enzyme
Y46G/S48G
positive cooperativity in binding the coenzyme NAD+
Y46G/S48G
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inactive mutant enzyme, only exists as dimer
D311N
site-directed mutagenesis, the mutation breaks the salt bridge between the catalytic and NAD+-binding domains, mutant dN-GAPDS D311N binds NAD+ noncooperatively
D311N
site-directed mutagenesis, the mutation breaks the salt bridge between the catalytic and NAD+-binding domains, the inactivation rate constant in the presence of GdnHCl increases 6fold, and the value of GdnHCl concentration corresponding to the protein half-denaturation decreases from 1.83 to 1.35 M. The mutation D311N enhances the enzymatic activity of the protein 2fold
E244Q
site-directed mutagenesis, mutation at the interdomain salt bridge
E244Q
site-directed mutagenesis, mutation at the interdomain salt bridge, the E244Q substitution does not alter the NAD+-binding significantly. The mutant protein exhibits a well-pronounced positive cooperativity in coenzyme binding
E96Q
site-directed mutagenesis, mutation at the interdomain salt bridge
E96Q
site-directed mutagenesis, mutation at the interdomain salt bridge, the E96Q substitution does not alter the NAD+-binding significantly. The mutant protein exhibits a well-pronounced positive cooperativity in coenzyme binding
C150G
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the Tdh3 mutation eliminates catalytic activity, the mutant is defective in silencing and deficient in NAD+ binding
C150G
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the Tdh3 mutation eliminates catalytic activity, the mutant is defective in silencing and deficient in NAD+ binding
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additional information
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recombinant expression of Gapdh from Thermoanaerobacterium saccharolyticum in Clostridium thermocellum improves the growth rates of the cells in presence of ethanol compared to wild-type
additional information
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GAPC-1 antisense line shows a delay in growth, morpholigical alterations in siliques, and low seed number. Embryo development is altered, showing abortions and empty embryonic sacs in basal and apical siliques, respectively. Mutant shows a decrease in the expression and activity of aconitase and succinate dehydrogenase and reduced levels of pyruvate and several Krebs cycle intermediates, and increased reactive oxygen species levels
additional information
construction of enzyme gapcp double mutants, gapcp1gapcp2, under the control of photosynthetic (Rubisco small subunit RBCS2B [RBCS]) or heterotrophic (phosphate transporter PHT1.2 [PHT]) cell-specific promoters
additional information
construction of enzyme gapcp double mutants, gapcp1gapcp2, under the control of photosynthetic (Rubisco small subunit RBCS2B [RBCS]) or heterotrophic (phosphate transporter PHT1.2 [PHT]) cell-specific promoters
additional information
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construction of glyceraldehyde-3-phosphate dehydrogenase double mutant gapcp1gapcp2. GAPCp expression in photosynthetic cells of gapcp1-gapcp2 does not complement the growth arrest of the aerial parts of the mutant plants, the lack of GAPCp activity in epidermal cells restricts leaf growth
additional information
knockout or overexpression of GAPC isozymes in Arabidopsis thaliana causing significant changes in the level of intermediates in the glycolytic pathway and the ratios of ATP/ADP and NAD(P)H/NAD(P). Construction of two double knockout seeds by T-DNA insertion showing over 3% of dry weight decrease in oil content compared with that of the wild-type. In transgenic seeds under the constitutive 35S promoter, oil content is increased up to 42% of dry weight compared with 36% in the wild-type and the fatty acid composition is altered. The transgenic lines exhibit decreased fertility. Seed-specific overexpression lines show over 3% increase in seed oil without compromised seed yield or fecundity, phenotypes, overview
additional information
knockout or overexpression of GAPC isozymes in Arabidopsis thaliana causing significant changes in the level of intermediates in the glycolytic pathway and the ratios of ATP/ADP and NAD(P)H/NAD(P). Construction of two double knockout seeds by T-DNA insertion showing over 3% of dry weight decrease in oil content compared with that of the wild-type. In transgenic seeds under the constitutive 35S promoter, oil content is increased up to 42% of dry weight compared with 36% in the wild-type and the fatty acid composition is altered. The transgenic lines exhibit decreased fertility. Seed-specific overexpression lines show over 3% increase in seed oil without compromised seed yield or fecundity, phenotypes, overview
additional information
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knockout or overexpression of GAPC isozymes in Arabidopsis thaliana causing significant changes in the level of intermediates in the glycolytic pathway and the ratios of ATP/ADP and NAD(P)H/NAD(P). Construction of two double knockout seeds by T-DNA insertion showing over 3% of dry weight decrease in oil content compared with that of the wild-type. In transgenic seeds under the constitutive 35S promoter, oil content is increased up to 42% of dry weight compared with 36% in the wild-type and the fatty acid composition is altered. The transgenic lines exhibit decreased fertility. Seed-specific overexpression lines show over 3% increase in seed oil without compromised seed yield or fecundity, phenotypes, overview
additional information
generation of a gapc1/gapc2 double mutant that is entirely devoid of the cytosolic GAPC activity and insensitive to Tyr-Asp inhibition of GAPC activity
additional information
generation of a gapc1/gapc2 double mutant that is entirely devoid of the cytosolic GAPC activity and insensitive to Tyr-Asp inhibition of GAPC activity
additional information
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knockout or overexpression of GAPC isozymes in Arabidopsis thaliana causing significant changes in the level of intermediates in the glycolytic pathway and the ratios of ATP/ADP and NAD(P)H/NAD(P). Construction of two double knockout seeds by T-DNA insertion showing over 3% of dry weight decrease in oil content compared with that of the wild-type. In transgenic seeds under the constitutive 35S promoter, oil content is increased up to 42% of dry weight compared with 36% in the wild-type and the fatty acid composition is altered. The transgenic lines exhibit decreased fertility. Seed-specific overexpression lines show over 3% increase in seed oil without compromised seed yield or fecundity, phenotypes, overview
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additional information
the coenzyme specificity of GAPDH, EC 1.2.1.12, of Corynebacterium glutamicum is systematically manipulated by rational protein design and the effect of the manipulation for cellular metabolism and lysine production is evaluated. By a combinatorial modification of four key residues within the coenzyme binding sites, different GAPDH mutants with varied coenzyme specificity are constructed. While increasing the catalytic efficiency of GAPDH towards NADP+ enhances lysine production in all of the tested mutants, the most significant improvement of lysine production (about 60%) is achieved with the mutant showing similar preference towards both NAD+ and NADP+, EC 1.2.1.59
additional information
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the coenzyme specificity of GAPDH, EC 1.2.1.12, of Corynebacterium glutamicum is systematically manipulated by rational protein design and the effect of the manipulation for cellular metabolism and lysine production is evaluated. By a combinatorial modification of four key residues within the coenzyme binding sites, different GAPDH mutants with varied coenzyme specificity are constructed. While increasing the catalytic efficiency of GAPDH towards NADP+ enhances lysine production in all of the tested mutants, the most significant improvement of lysine production (about 60%) is achieved with the mutant showing similar preference towards both NAD+ and NADP+, EC 1.2.1.59
additional information
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replacement of Escherichia coli GapA glyceraldehyde 3-phosphate dehydrogenase by Clostridium acetobutylicum GapC glyceraldehyde 3-phosphate dehydrogenase, EC 1.2.1.9 results in significant reduction of flux through the pentose phosphate pathway. Recombinant strains display increased NADPH availability, and consistently higher productivity than parent strains
additional information
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construction of a gapA-deficient strain MC4100 DELTAgapA
additional information
the gapA gene from Escherichia coli strain MG1655 is replaced by the gene gapN from Streptococcus mutans, EC 1.2.1.9, UniProt ID Q59931. The specific NADP+-GAPDH activity of the strain MG1655DgapA::gapN is 4.6times lower relative to strain MG1655DELTAgapA::gapN/pTrcgapN and no NAD+-GAPDH activity is detected. The specific NADP+-GAPDH activity levels in the derivative strain reveal that growth rate and glucose uptake differences are attributable to gapN expression level. The NADH/NAD+ ratio in the strain MG1655DELTAgapA::gapN/pTrcgapN decreases by 25% as compared to wild-type strain. In contrast, the NADPH/NADP+ ratio increases 2times indicating that the alteration in the turnover of NAD(P)H via glyceraldehyde 3-phosphate oxidation affects the redox levels of the strain MG1655DELTAgapA::gapN/pTrcgapN, which increases 2.8times the NADPH/NADH ratio
additional information
generation of an engineered synthetic Escherichia coli codon optimized sequence of a human gene that codifies for a 32.4 kDa protein for recombinant expression
additional information
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mutant htt shows co-localization of GAPDH with N-terminus of huntingtin aggregates
additional information
expression of a highly soluble form of GAPDS truncated at the N-terminus, amino acids 69398. Mutant displays a 3fold increase in catalytic efficiency and shows homotetrameric structure
additional information
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expression of a highly soluble form of GAPDS truncated at the N-terminus, amino acids 69398. Mutant displays a 3fold increase in catalytic efficiency and shows homotetrameric structure
additional information
construction of a plasmid encoding truncated GAPDS lacking 68 N-terminal amino acids (dN-GAPDS)
additional information
construction of a plasmid encoding truncated GAPDS lacking 68 N-terminal amino acids (dN-GAPDS)
additional information
construction of a plasmid encoding truncated GAPDS lacking 68 N-terminal amino acids (dN-GAPDS). The recombinant GAPDS without the N-terminal sequence (dN-GAPDS) is soluble in contrast to the wild-type
additional information
construction of a plasmid encoding truncated GAPDS lacking 68 N-terminal amino acids (dN-GAPDS). The recombinant GAPDS without the N-terminal sequence (dN-GAPDS) is soluble in contrast to the wild-type
additional information
construction of three deletion mutants of GAPDH that lack different lengths in their C-terminal regions (GAPDH1-106, GAPDH1-176, and GAPDH1-230). Among these three mutants, GAPDH1-106 shows an affinity with SRR, whereas GAPDH1-176 and GAPDH1-230 do not, suggesting that the catalytic domain interrupts interacting sites in the NAD+-binding domain of GAPDH
additional information
viral mutations R58E, Q59A or Q63A in the matrix region, and E76R or R82E in the capsid N-terminal domain abrogate the interaction with the C-terminal domain of enzyme GAPDH. SAccharomyces cerevisiae two-hydrib interaction analysis between enzyme GAPDH wild-type and mutants with HIV-1 wild-type and mutant matrix region and capsid N-terminal domain, overview
additional information
knockdown of GAPDHS in uveal melanoma (UM) cell lines hinders glycolysis by decreasing glucose uptake, lactate production, ATP generation, cell growth and proliferation. Conversely, overexpression of GAPDHS promotes glycolysis, cell growth and proliferation. Transcription factor SOX10 knockdown reduces the activation of GAPDHS, leading to an attenuated malignant phenotype, and SOX10 overexpression promotes the activation of GAPDHS, leading to an enhanced malignant phenotype
additional information
GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation o the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate. MA-RGAPDH2 is found to strengthen the metabolic flux of dihydroxyacetone phosphate to glycerol-3-phosphate
additional information
GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation o the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate. MA-RGAPDH2 is found to strengthen the metabolic flux of dihydroxyacetone phosphate to glycerol-3-phosphate
additional information
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GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation o the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate. MA-RGAPDH2 is found to strengthen the metabolic flux of dihydroxyacetone phosphate to glycerol-3-phosphate
additional information
GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation of the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate
additional information
GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation of the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate
additional information
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GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation of the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate
additional information
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GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation of the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate
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additional information
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GAPDH1 gene silencing using Agrobacterium tumefaciens strain AGL-1-mediated transformation o the cells with siRNA. RNA interference of isozymes GAPDH1 and GAPDH2 (MA-RGAPDH1 and MA-RGAPDH2) greatly reduced the biomass of the fungus. The lipid content of MA-RGAPDH2 is about 23% higher than that of the control. Both of the lipid-increasing transformants show a higher NADPH/NADP ratio. Analysis of metabolite and enzyme expression levels reveals that the increased lipid content of MA-GAPDH1 is due to enhanced flux of glyceraldehyde-3-phosphate to glycerate-1,3-biphosphate. MA-RGAPDH2 is found to strengthen the metabolic flux of dihydroxyacetone phosphate to glycerol-3-phosphate
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additional information
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mutants Gapdhm1Neu (Y91stop truncated protein of 89 amino acids), Gapdhm2Neu (truncated/altered protein of 9 amino acids), and Gapdhm8Neu (truncated/altered protein of 73 amino acids) exhibit 50-55% residual activity in blood compared to the wild type enzyme
additional information
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generation of FAO hepatoma cells with mutations of all 4 lysine residues K115, K160, K225, and K252 (4K-R-GAPDH) in critical regions of enzyme GAPDH to mimic their unmodified state reduces GAPDH glycolytic activity and glycolytic flux and increases gluconeogenic GAPDH activity and glucose production, phenotype overview
additional information
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generation of FAO hepatoma cells with mutations of all 4 lysine residues K115, K160, K225, and K252 (4K-R-GAPDH) in critical regions of enzyme GAPDH to mimic their unmodified state reduces GAPDH glycolytic activity and glycolytic flux and increases gluconeogenic GAPDH activity and glucose production, phenotype overview
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additional information
generation of a gapc1/gapc2 double mutant that is entirely devoid of the cytosolic GAPC activity and insensitive to Tyr-Asp inhibition of GAPC activity
additional information
generation of a gapc1/gapc2 double mutant that is entirely devoid of the cytosolic GAPC activity and insensitive to Tyr-Asp inhibition of GAPC activity
additional information
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a downregulation of GAPDH activity does not contribute to improved performance of engineered Saccharomyces cerevisiae on pentose substrates
additional information
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nuclear export sequences are fused to the 3' end of gene TDH3 by transforming a DNA fragment with 3' homology to the TDH3 ORF, the nuclear export sequence, and an hphMX4 sequence into the appropriate yeast strain. Strains lacking both TDH3 and specific NAD+ biosynthetic genes are generated by crossing Dtdh3 strain YSH969 with selected strains from the yeast deletion collection. Identification of sporulation haploid strains
additional information
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nuclear export sequences are fused to the 3' end of gene TDH3 by transforming a DNA fragment with 3' homology to the TDH3 ORF, the nuclear export sequence, and an hphMX4 sequence into the appropriate yeast strain. Strains lacking both TDH3 and specific NAD+ biosynthetic genes are generated by crossing Dtdh3 strain YSH969 with selected strains from the yeast deletion collection. Identification of sporulation haploid strains
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