5.3.1.1: triose-phosphate isomerase
This is an abbreviated version!
For detailed information about triose-phosphate isomerase, go to the full flat file.
Word Map on EC 5.3.1.1
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5.3.1.1
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giardia
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assemblage
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aldolase
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duodenalis
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dihydroxyacetone
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enolase
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phosphoglycerate
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zoonotic
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barrel
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fecal
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trypanosoma
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multilocus
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fructose
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giardiasis
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dhap
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cryptosporidium
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brucei
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protozoan
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isomerases
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gapdh
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province
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cysts
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malate
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stool
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lamblia
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methylglyoxal
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phosphofructokinase
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cruzi
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schistosoma
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enediolate
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2-phosphoglycolate
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parvum
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hemolytic
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intestinalis
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fructose-1,6-bisphosphate
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deamidation
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alpha-enolase
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1,6-bisphosphate
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glucosephosphate
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tim-barrel
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hungarian
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hominis
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phosphite
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dianion
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phosphoglucose
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glycosomes
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enzymopathy
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fructose-bisphosphate
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ige-binding
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subgenotype
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diagnostics
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synthesis
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analysis
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biofuel production
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medicine
- 5.3.1.1
- giardia
-
assemblage
- aldolase
- duodenalis
- dihydroxyacetone
- enolase
- phosphoglycerate
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zoonotic
-
barrel
-
fecal
- trypanosoma
-
multilocus
- fructose
- giardiasis
- dhap
- cryptosporidium
- brucei
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protozoan
- isomerases
- gapdh
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province
- cysts
- malate
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stool
- lamblia
- methylglyoxal
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phosphofructokinase
- cruzi
- schistosoma
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enediolate
- 2-phosphoglycolate
- parvum
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hemolytic
- intestinalis
- fructose-1,6-bisphosphate
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deamidation
- alpha-enolase
- 1,6-bisphosphate
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glucosephosphate
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tim-barrel
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hungarian
- hominis
- phosphite
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dianion
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phosphoglucose
- glycosomes
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enzymopathy
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fructose-bisphosphate
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ige-binding
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subgenotype
- diagnostics
- synthesis
- analysis
- biofuel production
- medicine
Reaction
Synonyms
CP 25, CTIMC, cTPI, cytoplasmic TPI, cytoplasmic triosephosphate isomerase, cytoTPI, D-glyceraldehyde-3-phosphate ketol-isomerase, GlTIM, Isomerase, triose phosphate, Lactacin B inducer protein, monoTIM, PfTIM, PfuTIM, Phosphotriose isomerase, plastidic TPI, plastidic triosephosphate isomerase, pTPI, SSO2592, TcTIM, TIM, TIM1, TIM2, TonTIM, TpI, TPI1, TpiA, Triose phosphate isomerase, Triose phosphate mutase, Triose phosphoisomerase, Triosephosphate isomerase, Triosephosphate mutase, vTIM
ECTree
5.3.1.1 triose-phosphate isomeraseAdvanced search results
results (
results (Engineering
Engineering on EC 5.3.1.1 - triose-phosphate isomerase
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PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
C218S
loss of about 21% of specific activity. In presence of glutathione disulfide, mutant behaves similarly to the wild-type, but retains more activity than the wild-type after 240 min of incubation
M80T
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analysis of key aspects of triosephosphate isomerase deficiency glycolytic enzymopathy pathogenesis identified using the TPIsugarkill mutation M80T, a Drosophila model of the human disease deficiency. Mutant protein is expressed, capable of forming a homodimer, and is functional. However, the mutant protein is degraded by the 20S proteasome core leading to loss-of-function pathogenesis
K174G/T175G/A176G
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16.5fold decrease in turnover number for D-glyceraldehyde 3-phosphate, 7.9fold increase in Km-value for D-glyceraldehyde 3-phosphate, 8fold decrease in turnover number for dihydroxyacetone phosphate, 2.6fold increase in Ki-value for arsenate, 10.4fold increase in Ki-value for 2-phosphoglycolate
L7RM
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the mutant exhibits a 200fold decrease in kcat/Km for isomerization of D-glyceraldehyde 3-phosphate and dihydroxyacetone phosphate. The mutant exhibits a 25fold decrease in kcat/Km for deprotonation of glycolaldehyde catalyzed by free enzyme. The mutation has little effect on the observed and intrinsic phosphodianion binding energy and only a modest effect on phosphite dianion activation of the enzyme
V167G/W168G
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17.9fold decrease in turnover number for D-glyceraldehyde 3-phosphate, 2.6fold increase in Km-value for D-glyceraldehyde 3-phosphate, 8.6fold decrease in turnover number for dihydroxyacetone phosphate, 2.7fold increase in Ki-value for arsenate, 8.1fold increase in Ki-value for 2-phosphoglycolate
V167G/W168G/K174G/T175G/A176G
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2529fold decrease in turnover number for D-glyceraldehyde 3-phosphate, 8.5fold increase in Km-value for D-glyceraldehyde 3-phosphate, 1720fold decrease in turnover number for dihydroxyacetone phosphate, 2.8fold increase in Ki-value for arsenate, 12.4fold increase in Ki-value for 2-phosphoglycolate
V167P/W168E
loop replacement mutant, 23000fold decrease in kcat/Km value. Mutations result in large displacement of the side chain of E168 from that for W168 in wild-type. Binding of glycerol 3-phosphate results in chemical shift changes for nuclei at the active site that are smaller than those of wild-type
H12N
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mutant enzyme with decreased thermal stability compared to wild-type enzyme. Half-life of 11.5 min at 64°C compared to 68°C for the wild-type enzyme
H12N/K13G
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mutant enzyme with decreased thermal stability compared to wild-type enzyme. Half-life of 11.5 min at 37°C compared to 68°C for the wild-type enzyme
H13G
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mutant enzyme with decreased thermal stability compared to wild-type enzyme. Half-life of 11.5 min at 51°C compared to 68°C for the wild-type enzyme
F12W
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W162F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W162F/W173F/W196F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W173F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W196F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W75F
mutant constructed for fluorescence quenching studies, catalytic properties similar to wild-type
W75F/W162F/W173F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W75F/W162F/W196F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
W75F/W173F/W196F
mutant constructed for fluorescence studies, catalytic properties similar to wild-type
D213A
D213Q
K183A
K183S
E104D
I170V
E65Q
the variant of Leishmania mexicana TIM has a much enhanced stability but its catalytic properties are the same as wild-type leishmanial TIM
A238S
C126A
the mutant shows an approximately 5.8fold drop in kcat compared to the wild type enzyme
C126M
the mutant shows an approximately 10fold drop in catalytic activity compared to the wild type enzyme
C126S
the mutant shows an approximately 5.8fold drop in kcat compared to the wild type enzyme
C126T
the mutant shows an approximately 10fold drop in catalytic activity compared to the wild type enzyme
C126V
the mutant shows an approximately 10fold drop in catalytic activity compared to the wild type enzyme
C13D
the mutant displays significant reduction in catalytic activity when compared with wild type enzyme (about 7.4fold decrease in kcat). The C13D mutant dissociates at concentrations above 1.25 mM
C13E
the mutant displays significant reduction in catalytic activity when compared with wild type enzyme (about 3.3fold decrease in kcat). The C13E mutant retains dimeric at concentrations above 1.25 mM
E165A
the mutant shows an approximately 9000fold drop in activity
F96H
site-directed mutagenesis, the mutant exhibits highly reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme
F96S
F96W
site-directed mutagenesis, the mutant exhibits reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme. The wild-type enzyme shows a loop-open state for 3-phosphoglycerate binding at the active site, while the mutant F96W shows both open and closed states
Q64E
Q64N
T75C
activity similar to wild-type, dimer integrity is unimpaired. Decrease in stability between 35°C and 45°C
T75N
4fold drop in activity compared to wild-type. Decrease in stability between 35°C and 45°C
T75S
T75V
activity similar to wild-type, dimer integrity is unimpaired. Decrease in stability between 35°C and 45°C
W11F/W168F/Y74W
site-directed mutagenesis, template is the available crystal structure of the enzyme from Giardia lamblia, which contains a Trp residue at position 47. The mutant dissociates at low protein concentrations, and exhibits considerably reduced stability in the presence of denaturants, urea and guanidinium chloride, and it shows approximately 20fold reduction in kcat at low protein concetrations compared to the wild-type enzyme, but the mutant mutant shows an enhancement of activity of 21.9fold at higher concentration range
Y74C
Y74G
site-directed mutagenesis, the mutation Tyr74Gly significantly reduces the stability of the dimer, mutation-induced alteration in the backbone conformation of Lys12, structure comparison to the wild-type enzyme
R111T/R112A/E114K/E115G
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site-directed mutagenesis, destroying of four ion pair interactions by replacing four of the charged residues in helix 4 with structurally analogous residues from the psychrophile Methanococcoides burtonii TIM to create a mutant, mPfuTIM, that is less kinetically stable than wild-type PfuTIM
C126A
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turnover number for D-glyceraldehyde 3-phosphate is 1.5fold lower than the wild-type value, KM-value for D-glyceraldehyde 3-phosphate is 1.4fold lower than the wild-type value, turnover number for dihydroxyacetone phosphate is 4.3fold lower than the wild-type value, KM-value for dihydroxyacetone phosphate is 3.7fold lower than the wild-type value
C126S
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mutant enzyme shows greater susceptibility to thermal denaturation than wild-type enzyme, turnover number for dihydroxyacetone phosphate is 8.3fold lower than the wild-type value, KM-value for dihydroxyacetone phosphate is 3.5fold lower than the wild-type value
D225Q
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mutation causes minor drops in Km and kcat value without changes catalytic efficiency. Temperature-induced unfolding-refolding of both wild-type and mutant D225Q samples display hysteresis cycles, indicative of processes far from equilibrium. The rate constant for unfolding is about three-fold larger in the mutant than in wild-type. Upon mutation, the rate-limiting step changes from a second-order at submicromolar concentrations to a first-order reaction. Renaturation occurs through a uni-bimolecular mechanism in which refolding of the monomer most likely begins at the C-terminal half of its polypeptide chain
I170A
effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
I170A/L230A
effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
K12G
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the mutation results in a ca. 50fold increase in Km for the substrate glyceraldehyde 3-phosphate (GAP) and a 60fold increase in Ki for competitive inhibition by 2-phosphoglycolate, a 12000fold decrease in kcat for isomerization of GAP, and a 6000000fold decrease in kcat/Km for GAP
K17A/Y46A/D48F/Q82A/D85S
mutation of residues in the dimer interface of enzyme
K17L/Y46F/D48F/Q82F/D85L
mutation of residues in the dimer interface of enzyme. Decrease in catalytic efficiency by 4 orders of magnitude
K17L/Y46F/D48Y/Q82A/D85A
mutation of residues in the dimer interface of enzyme. Decrease in catalytic efficiency by 4 orders of magnitude
K17P/Y46A/D48L/Q82T/D85A
mutation of residues in the dimer interface of enzyme
L230A
effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
S211A
mutation eliminates intraloop hydrogen bonds to the side-chain hydroxyl, 60fold decrease in kcat/Km
S211G
mutation eliminates intraloop hydrogen bonds to the side-chain hydroxyl, leading to small changes in the kinetic parameters
Y208A
main effect of mutations is to cause a reduction in the total intrinsic dianion binding energy
Y208F
mutation eliminates the intraloop hydrogen bond between the hydroxyl group of Y208 and the amide nitrogen of A176. Enzyme activity is reduced by ca. 50fold compared to the Y208A and Y208S mutants
Y208S
main effect of mutations is to cause a reduction in the total intrinsic dianion binding energy
Y208T
main effect of mutations is to cause a reduction in the total intrinsic dianion binding energy
I170A
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effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
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I170A/L230A
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effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
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L230A
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effect of the mutation on the relative electrostatic contribution of the residue is negligible. Mutation results in increases in the activation barriers for deprotonation of substrate
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H12N/K13G
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mutant enzyme shows increases thermal stability compared to wild-type enzyme. Half-life of 11.5 min at 96°C compared to 94°C of the wild-type enzyme
I45A
V45A
29fold less active than wild-type. Mutant dissociates into stable monomers and assembles as catalytic competent dimer upon binding of the transition state substrate analog phosphoglycolohydroxamate
A178L
decrease in catalytic efficiency. Crystallization data reveal a more disordered loop-6 in the structure of unliganded A178L. Liganded structures show minimal differences to wild-type
C14A
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the KM-value for D-glyceraldehyde 3-phosphate is 1.2fold higher than the value for the wild-type enzyme. The KM-value for glycerone phosphate is 90% of the value for the wild-type enzyme. The turnover number for D-glyceraldehyde 3-phosphate is 1.2fold higher than the value for the wild-type enzyme. The turnover-number for glycerone phosphate is comparable to the value for the wild-type enzyme
C14F
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the Ki-value for 3-phosphoglycolate is nearly 3times higher than the Ki-value for the wild-type enzyme. The ratio of elimination to isomerization reactions is higher than in the wild-type enzyme. The KM-value for D-glyceraldehyde 3-phosphate is 10.2fold higher than the value for the wild-type enzyme. The KM-value for glycerone phosphate is 2.7fold higher than the value for the wild-type enzyme. The turnover number for D-glyceraldehyde 3-phosphate is 2280fold lower than the value for the wild-type enzyme. The turnover-number for glycerone phosphate is 3000fold lower than the value for the wild-type enzyme
C14L
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Cys14Leu mutant has the tendency to aggregate, reduced stability and altered kinetics
C14P
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the KM-value for D-glyceraldehyde 3-phosphate is 94% of the value for the wild-type enzyme. The KM-value for glycerone phosphate is 58% of the value for the wild-type enzyme. The turnover number for D-glyceraldehyde 3-phosphate is 85% of the value for the wild-type enzyme. The turnover-number for glycerone phosphate is 81% of the value for the wild-type enzyme
C14S
C14S/A69C
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similar in kinetic parameters to wild-type TbTIM and the single mutant C14S. Mutant binds 50 times more 1-anilino-8-naphthalene sulfonate than wild-type and is susceptible to digestion with subtilisin
C14S/A73C
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greatly reduced kcat value. Mutant binds 50 times more 1-anilino-8-naphthalene sulfonate than wild-type and is susceptible to digestion with subtilisin
C14S/S71C
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similar in kinetic parameters to wild-type TbTIM and the single mutant C14S. Mutant binds 50 times more 1-anilino-8-naphthalene sulfonate than wild-type and is susceptible to digestion with subtilisin
C14S/S79C
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similar in kinetic parameters to wild-type TbTIM and the single mutant C14S. Mutant binds 50 times more 1-anilino-8-naphthalene sulfonate than wild-type and is susceptible to digestion with subtilisin
C14T
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the KM-value for D-glyceraldehyde 3-phosphate is 1.2fold higher than the value for the wild-type enzyme. The KM-value for glycerone phosphate is 79% of the value of the wild-type enzyme. The turnover number for D-glyceraldehyde 3-phosphate is 92% of the value for the wild-type enzyme. The turnover-number for glycerone phosphate is 92% of the value of the wild-type enzyme
D227A
mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2.2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 5fold lower than the value for the wild-type enzyme
D227N
mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 3.75fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 3.5fold lower than the value for the wild-type enzyme
E168D
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catalytically inert. Dimer formation with Trypanosoma cruzi dimer interface mutant C15Acauses a drop in activity by 50%
H47N
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mutant His47Asn, dimer is considerably less stable than wild-type trypanosomal enzyme
P168A
mutation beside catalytic residue Glu167. Mutant turnover number is 50fold reduced, Km value is 2fold reduced
R191A
mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 10.4fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 10.6fold lower than the value for the wild-type enzyme
R191S
mutant enzyme with reduced stability to 3.2 M guanidinium-HCl and reduced thermal stability at 42°C. The ratio of turnover number to Km for D-glyceraldehyde 3-phosphate as substrate is 2fold lower than the value for the wild-type enzyme. The ratio of turnover number to Km for glycerone phosphate as substrate is 6.7fold lower than the value for the wild-type enzyme
E23G/A70T/S96F/A178V
mutant facilitates better growth of a Escherichia coli L-arabinose isomerase knockout strain in medium supplemented with 40 mM L-arabinose. Mutant shows increased thermostability
F28L/A100L/A115Q
mutant constructed to mimick the methylmethane sulfonate inactivation pattern of the Trypanosoma cruzi enzyme. Protein is similarly susceptibel to methylmethane sulfonate as the Trypanosoma cruzi enzyme
L232A
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the mutation in a 6fold decrease in kcat/Km for the reversible isomerization of D-glyceraldehyde 3-phosphate to give dihydroxyacetone phosphate
P168A
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the mutation results in 50fold decrease in kcat/Km for deprotonation of glycolaldehyde catalyzed by free enzyme
Q65L
mutant facilitates better growth of a Escherichia coli L-arabinose isomerase knockout strain in medium supplemented with 40 mM L-arabinose. Mutant shows increased thermostability
S43P
mutation increases the free energy of the transition state by 17.7 kJ/mol
C15A
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dimer formation of mutant protein with intact Trypanosoma brucei monomer, maximal inhibition of catalysis by mutation is 60%. Dimer formation with catalytically inert Trypanosoma brucei mutant E168D causes a drop in activity by 50%
E26D/T27L/L28F/A30S/L100A/Q115A
mutant construcuted to produce a TIM with the inactivation susceptibility pattern of Trypanosoma brucei brucei enzyme, shows a similar resistance pattern to the inactivation with methylmethane thiosulfonate as wild-type Trypanosoma brucei brucei TIM
E26D/T27L/L28F/A30S/T32S/L100A/Q115A
mutant construcuted to produce a TIM with the inactivation susceptibility pattern of Trypanosoma brucei brucei enzyme, shows a similar resistance pattern to the inactivation with methylmethane thiosulfonate as wild-type Trypanosoma brucei brucei TIM
E26D/T27L/L28F/L100A/Q115A
mutant construcuted to produce a TIM with the inactivation susceptibility pattern of Trypanosoma brucei brucei enzyme, shows a similar resistance pattern to the inactivation with methylmethane thiosulfonate as wild-type Trypanosoma brucei brucei TIM
L28F/L100A/Q115A
mutant construcuted to produce a TIM with the inactivation susceptibility pattern of Trypanosoma brucei brucei enzyme. Protein is more resistant to the inactivation with methylmethane thiosulfonate as wild-type
additional information
D213A
Residue putatively involved in a highly conserved salt bridge lacking in Helicobacter pylori enzyme. Kinetics and isomerization activity similar to wild-type
D213A
disruption of the salt bridge between residues D213 and K183, kinetic parameters similar to wild-type
D213Q
Residue putatively involved in a highly conserved salt bridge lacking in Helicobacter pylori enzyme. Kinetics and isomerization activity similar to wild-type
D213Q
disruption of the salt bridge between residues D213 and K183, kinetic parameters similar to wild-type
K183A
Residue putatively involved in a highly conserved salt bridge lacking in Helicobacter pylori enzyme. Kinetics and isomerization activity similar to wild-type
K183A
disruption of the salt bridge between residues D213 and K183, kinetic parameters similar to wild-type
K183S
Residue putatively involved in a highly conserved salt bridge lacking in Helicobacter pylori enzyme. Kinetics and isomerization activity similar to wild-type
K183S
disruption of the salt bridge between residues D213 and K183, kinetic parameters similar to wild-type
E104D
mutation involved in human triosephosphate isomerase defiecency autosomal disease. Mutant exhibits normal catalytic activity but shows impairments in the formation of active dimers and low thermostability. E104A is part of a conserved cluster of 10 residues, 5 from each subunit. This cluster forms a cavity that possesses an elaborate conserved network of buried water molecules that bridge the two subunits. In the E104D mutant, a disruption of contacts of the amino acid side chains in the conserved cluster leads to a perturbation of the water network in which the water-protein and water-water interactions that join the two monomers are significantly weakened and diminished
E104D
in patients homozygous for this mutation, only 2-20% of TIM catalytic activity is left
A238S
-
mutant enzyme A238S has a higher thermal stability than the wild-type enzyme. The turnover number of the mutant enzyme is somewhat lower than that observed for the wild-type enzyme, the Km-value for D-glyceraldehyde 2-phosphate is somewhat higher
A238S
-
catalytic efficiency of the mutant enzyme is somewhat reduced, and its stability is considerably increased. The half-life at 25°C is 27 min compared to 10 min for the wild-type enzyme
F96S
site-directed mutagenesis, the mutant exhibits highly reduced catalytic efficiency and decreased substrate-binding affinity, as well as reduced sensitivity to inhibitor 3-phosphoglycerate, compared to the wild-type enzyme
Q64E
mutation at key dimer interface residue, decreases dimer stability
Q64N
mutation at key dimer interface residue, decreases dimer stability
T75S
activity similar to wild-type, dimer integrity is unimpaired. Decrease in stability between 35°C and 45°C
T75S
mutation at key dimer interface residue, decreases dimer stability
Y74C
inhibitory effect of KFGNGSYTGEVS and KYGNGSCTGEVS is more pronounced compared to the wild-type enzym. Mutant enzyme has reduced stability due to an interface cavity
Y74C
-
urea unfolding profiles of Y74Cox in urea solution obtained by fluorescence and circular dichroism approximate a two-state transition and do not show the presence of stable intermediates over a wide range of denaturant concentrations. In wild-type enzyme the unfolding results in gradual change in spectroscopic properties. The unfolding transition midpoit is 3.5 M for Y74Cox and 5.5 M for the wild-type enzyme
I45A
29fold decrease in activity. Residue I45 controls the dimer-monomer equilibrium, mutant displays a dimer-monomer equilibrium and is stable
I45A
39fold less active than wild-type. Mutant dissociates into stable monomers and assembles as catalytic competent dimer upon binding of the transition state substrate analog phosphoglycolohydroxamate
C14S
-
the KM-value for D-glyceraldehyde 3-phosphate is 1.4fold higher than the value for the wild-type enzyme. The KM-value for glycerone phosphate is identical to the value of the wild-type value. The turnover number for D-glyceraldehyde 3-phosphate is 1.15fold higher than the value for the wild-type enzyme. The turnover-number for glycerone phosphate is nearly identical to the value of the wild-type enzyme
C14S
-
wild-type-like enzyme that is resistant to the action of sulfhydryl reagents methylmethane thiosulfonate and 5,5-dithiobis(2-nitrobenzoate)
additional information
construction of a T-DNA insertion mutant that shows a fivefold reduction in transcript, reduced TPI activity, and a severely stunted and chlorotic seedling that accumulates dihydroxyacetone phosphate, glycerol, and glycerol-3-phosphate. Methylglyoxal, a by-product of dihydroxyacetone phosphate, also accumulated in the pdtpi mutant. Lipid profiling reveals lower monogalactosyl but higher digalactosyl lipids in the mutant compared to the wild-type, phenotype, overview. Exogenous glycolytic intermediates do not rescue the pdtpi phenotype. Metabolic pathways affected by a mutation in pdTPI during postgerminative seedling transition from heterotrophic to photoautotrophic development, overview
additional information
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construction of a T-DNA insertion mutant that shows a fivefold reduction in transcript, reduced TPI activity, and a severely stunted and chlorotic seedling that accumulates dihydroxyacetone phosphate, glycerol, and glycerol-3-phosphate. Methylglyoxal, a by-product of dihydroxyacetone phosphate, also accumulated in the pdtpi mutant. Lipid profiling reveals lower monogalactosyl but higher digalactosyl lipids in the mutant compared to the wild-type, phenotype, overview. Exogenous glycolytic intermediates do not rescue the pdtpi phenotype. Metabolic pathways affected by a mutation in pdTPI during postgerminative seedling transition from heterotrophic to photoautotrophic development, overview
additional information
-
mutation wasted away is a recessive, hypomorphic mutation that causes progressive motor impairment, vacuolar neuropthology, and severely reduced lifespan. Mutation affects the gene for triosephosphate isomerase. There is no genetic evidence that the mutation leads to misfolded or aberrant protein. Mutation may lead to an accumulation of methylglyoxal and the consequent enhanced production of advanced glycation end products
additional information
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overexpression of isoform Tpi2 in a Tpi1-deficient mutant restores the growth deficiency on minimal medium containing glucose or glycerol
additional information
overexpression of isoform Tpi2 in a Tpi1-deficient mutant restores the growth deficiency on minimal medium containing glucose or glycerol
additional information
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Tpi1-deficient mutant grows weakly on minimal medium containing glucose or glycerol, but grows normally on gluconate. Residual enzymic activity on glucose or glycerol is due to presence of enzyme isoform Tpi2
additional information
Tpi1-deficient mutant grows weakly on minimal medium containing glucose or glycerol, but grows normally on gluconate. Residual enzymic activity on glucose or glycerol is due to presence of enzyme isoform Tpi2
additional information
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modulation of gene expression around wild-type level by replacing the native promoter with libraries of synthetic promoters. Enzyme is present in high excess in wild-type cells. 10% residual activity still support more than 70% of the wild-type glycolytic flux. At at residual triosephosphate isomerase activity of 3%, dihydroxyacetone phosphate level increases four times and coincides with an increase in formate production
additional information
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some TPIs are found on the paired helical filaments in Tau transgenic mice
additional information
decrease in ligand affinity in F96 mutants can be a consequence of differences in the water network connecting residue 96 to Ser73 in the vicinity of the active site
additional information
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decrease in ligand affinity in F96 mutants can be a consequence of differences in the water network connecting residue 96 to Ser73 in the vicinity of the active site
additional information
expression of enzyme confers arsenic tolerance to Escherichia coli XL1 Blue and DF502
additional information
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expression of enzyme confers arsenic tolerance to Escherichia coli XL1 Blue and DF502
additional information
mutation of five residues in the dimer interface of enzyme. Obtained proteins are soluble, dimeric, and compact. Proteins obtained from direct evolution experiments show wild-type-like catalytic activity, while their stability is decreased. In silico-designed proteins are very stable dimers that bind substrate with a wild-type-like Km value, albeit they exhibit a very low kcat
additional information
-
mutation of five residues in the dimer interface of enzyme. Obtained proteins are soluble, dimeric, and compact. Proteins obtained from direct evolution experiments show wild-type-like catalytic activity, while their stability is decreased. In silico-designed proteins are very stable dimers that bind substrate with a wild-type-like Km value, albeit they exhibit a very low kcat
additional information
a mutant lacking loop 3 is 1500fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
additional information
a mutant lacking loop 3 is 1500fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
additional information
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a mutant lacking loop 3 is 1500fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
additional information
a mutant lacking loop 3 is 9400fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
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a mutant lacking loop 3 is 9400fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
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a mutant lacking loop 3 is 9400fold less active than wild-type and dissociates into stable monomers. Upon binding of the transition state substrate analog phosphoglycolohydroxamate, mutant remains monomeric
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wild-type enzyme consists of two identical subunits that form a very tight dimer involving interactions of 32 residues of each subunit. By replacing 15 residues of the major interface loop by another 8-residue fragment, a variant is constructed that is a stable and monomeric protein with TIM activity, monoTIM. The turnover numer of the monomeric form is 100fold lower
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hybrids of Trypanosoma cruzi enzyme carrying residues of the dimer interface from Trypanomsoma brucei and vice versa, and hybrids with one monomer in the enzyme dimer from Trypanosoma cruzi, and the second monomer from Trypanosoma brucei. Solvent exposure of the interfacial C15 depends predominantly on the characteristics of the adjoining monomer. Half of the activity of each monomer depends on the integrity of each of the two C15-loop3 portions of the interface
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introduction of mutation V233A into a monomeric enzyme variant with an engineered binding groove, m18bTIM. Mutation V233A restores the structural properties of loop-7, the binding site of a conserved water molecule between loop-7 and loop-8 and the binding site of the phosphate moiety of wild-type in the m18bTIM background. The active site of the V233A mutant can bind transition state analogs and suicide inhibitors competently, the catalytic efficiency of the V233A mutant is too low to be detected
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a reporter protein fused to a Saccharomyces cerevisiae peptide containing the sequence corresponding to the 22-residue fragment, including residues K155, D158, W159, A160 and K161, of the Trypanosoma brucei TPI enzyme, is not targeted to glycosomes
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the monomeric form of TIM does not catalyze any reactions of D-glyceraldehyde 3-phosphate at the enzyme active site, either in the absence or in the presence of phosphite dianion
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analysis of chimera between Trypanosoma cruzi and Trypanosoma brucei brucei enzymes. The (betaalpha)2 motif of Trypanosoma cruzi TIM inserted into Trypanosoma brucei brucei TIM increases the kinetic stability. The presence of the (betaalpha)2 motif of Trypanosoma brucei brucei TIM inserted into Trypanosoma cruzi TIM gives a chimerical protein difficult to purify in soluble form and with a significantly reduced kinetic stability
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construction of mutants for interconversion of the behavior of the enzymes from Trypanosoma brucei brucei and Trypanosoma cruzi. To make Trypanosoma cruzi TIM reactivate with a Trypanosoma brucei brucei TIM-like behaviour you need to mutate the following amino acids: 18, 19, 20, 22, 23 and 26, and R2: 43, 46, 48, 49, 53, 56 and 57. To make a Trypanosoma brucei brucei TIM reactivate with a Trypanosoma cruzi TIM-like behaviour you need to mutate amino acids 18, 23, 26, 32, 33 and 34, and 43, 46, 48, 49, 53, 56 and 57
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hybrids of Trypanosoma cruzi enzyme carrying residues of the dimer interface from Trypanomsoma brucei and vice versa, and hybrids with one monomer in the enzyme dimer from Trypanosoma cruzi, and the second monomer from Trypanosoma brucei. Solvent exposure of the interfacial C15 depends predominantly on the characteristics of the adjoining monomer. Half of the activity of each monomer depends on the integrity of each of the two C15-loop3 portions of the interface
additional information
analysis of chimera between Trypanosoma cruzi and Trypanosoma brucei brucei enzymes. The (betaalpha)2 motif of Trypanosoma cruzi TIM inserted into Trypanosoma brucei brucei TIM increases the kinetic stability. The presence of the (betaalpha)2 motif of Trypanosoma brucei brucei TIM inserted into Trypanosoma cruzi TIM gives a chimerical protein difficult to purify in soluble form and with a significantly reduces kinetic stability
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analysis of chimera between Trypanosoma cruzi and Trypanosoma brucei brucei enzymes. The (betaalpha)2 motif of Trypanosoma cruzi TIM inserted into Trypanosoma brucei brucei TIM increases the kinetic stability. The presence of the (betaalpha)2 motif of Trypanosoma brucei brucei TIM inserted into Trypanosoma cruzi TIM gives a chimerical protein difficult to purify in soluble form and with a significantly reduces kinetic stability
additional information
construction of mutants for interconversion of the behavior of the enzymes from Trypanosoma brucei brucei and Trypanosoma cruzi. To make Trypanosoma cruzi TIM reactivate with a Trypanosoma brucei brucei TIM-like behaviour you need to mutate the following amino acids: 18, 19, 20, 22, 23 and 26, and R2: 43, 46, 48, 49, 53, 56 and 57. To make a Trypanosoma brucei brucei TIM reactivate with a Trypanosoma cruzi TIM-like behaviour you need to mutate amino acids 18, 23, 26, 32, 33 and 34, and 43, 46, 48, 49, 53, 56 and 57
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construction of mutants for interconversion of the behavior of the enzymes from Trypanosoma brucei brucei and Trypanosoma cruzi. To make Trypanosoma cruzi TIM reactivate with a Trypanosoma brucei brucei TIM-like behaviour you need to mutate the following amino acids: 18, 19, 20, 22, 23 and 26, and R2: 43, 46, 48, 49, 53, 56 and 57. To make a Trypanosoma brucei brucei TIM reactivate with a Trypanosoma cruzi TIM-like behaviour you need to mutate amino acids 18, 23, 26, 32, 33 and 34, and 43, 46, 48, 49, 53, 56 and 57
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construction of a monomeric enzyme form with loop-1 one residue shorter than monoTIM, which is constructed by replacing 15 residues of the major interface loop by another 8-residue fragment. The catalytic activity and stability of the new seven-residue Loop-1 enzyme variant is similar with that of monoTIM
