3.5.1.B15: pyrazinamidase
This is an abbreviated version!
For detailed information about pyrazinamidase, go to the full flat file.
Word Map on EC 3.5.1.B15
-
3.5.1.B15
-
tuberculosis
-
mycobacterium
-
pzase
-
pyrazinoic
-
pza-resistant
-
bactec
-
pyrazinamide-resistant
-
enterocolitica
-
wayne
-
mdr-tb
-
middlebrooks
-
autoagglutination
-
pza-susceptible
-
kristensenii
-
kansasii
-
diagnostics
- 3.5.1.B15
- tuberculosis
- mycobacterium
- pzase
-
pyrazinoic
-
pza-resistant
-
bactec
-
pyrazinamide-resistant
- enterocolitica
-
wayne
-
mdr-tb
-
middlebrooks
-
autoagglutination
-
pza-susceptible
- kristensenii
- kansasii
- diagnostics
Reaction
Synonyms
ASAC_0847, BsPncA, More, nicotinamidase/pyrazinamidase, PH0999, PncA, PolyNic, pyrazinamidase, pyrazinamidase/nicotinamidase, PZAase, PZAse
ECTree
3.5.1.B15 pyrazinamidaseAdvanced search results
results (
results (Engineering
Engineering on EC 3.5.1.B15 - pyrazinamidase
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
top
PROTEIN VARIANTS 
ORGANISM
UNIPROT
COMMENTARY 
LITERATURE
A143T
-
site-directed mutagenesis, the mutation decreases the Km and kcat values of the enzyme
A143T/T168A/E173K
-
site-directed mutagenesis, the mutation decreases the Km and kcat values of the enzyme, the mutant shows reduced thermostability compared to wild-type
A146V
naturally occuring mutation, the mutant shows 72% reduced activity compared to wild-type, resistant strain
D126N
site-directed mutagenesis, the mutation causes pyrazinamide resistance, the mutation is located outside of active site and has an allosteric affect
D12A
D12G
D136G
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
D49N
E144K
naturally occuring mutation from PZA-resistant isolate, analysis of the resistance mechanism of the mutant strain
G78C
G97D
naturally occuring mutation, the mutant shows 90% reduced activity compared to wild-type, resistant strain
H51P
naturally occuring mutation, inactive mutant, resistant strain
H51R
H57D
I5T
naturally occuring mutation, the mutant shows 88% reduced activity compared to wild-type, resistant strain
K96R
naturally occurring mutation in the PncA catalytic region, binding cavity analysis shows an increase of 762.3 A3 in the volume of the mutant protein. Docking studies reveal that pyrazinamide (PZA) has a greater binding affinity for the wild-type protein in comparison to the mutant protein. The residues of flap region acquire more flexibility in mutant form of protein and thus move away from the active site. This leads to weak binding of the drug to the target residues. The mutation leads to a substantial increase in the binding cavity. This prohibits the enzyme from holding the drug properly and therefore pyrazinoic acid (PZA) cannot take its active form
L151S
-
site-directed mutagenesis, the mutant has a weakened binding affinity for pyrazinamide and reduced thermostability compared to the wild-type
L19R
naturally occuring mutation from PZA-resistant isolate
L85R
naturally occuring mutation, inactive mutant, resistant strain
N11K
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
P69T
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
R140H
naturally occuring mutation from PZA-resistant isolate
T135P
T142R
T47P
naturally occuring mutation, inactive mutant, resistant strain
T87M
site-directed mutagenesis, active mutant, susceptible strain
T92C
the naturally occuring mutation causes an increase in distance from metal ion position to enzyme active site, but it is considered as a polymorphism
V9A
naturally occuring mutation, the mutant shows 73% reduced activity compared to wild-type, resistant strain
V9G
naturally occuring mutation, the mutant shows 99% reduced activity compared to wild-type, resistant strain
D126N
-
site-directed mutagenesis, the mutation causes pyrazinamide resistance, the mutation is located outside of active site and has an allosteric affect
-
D12A
D12G
D49N
E144K
-
naturally occuring mutation from PZA-resistant isolate, analysis of the resistance mechanism of the mutant strain
-
G78C
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
-
K96R
-
naturally occurring mutation in the PncA catalytic region, binding cavity analysis shows an increase of 762.3 A3 in the volume of the mutant protein. Docking studies reveal that pyrazinamide (PZA) has a greater binding affinity for the wild-type protein in comparison to the mutant protein. The residues of flap region acquire more flexibility in mutant form of protein and thus move away from the active site. This leads to weak binding of the drug to the target residues. The mutation leads to a substantial increase in the binding cavity. This prohibits the enzyme from holding the drug properly and therefore pyrazinoic acid (PZA) cannot take its active form
-
L19R
-
naturally occuring mutation from PZA-resistant isolate
-
N11K
-
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
-
P69T
-
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
-
R140H
-
naturally occuring mutation from PZA-resistant isolate
-
T92C
-
the naturally occuring mutation causes an increase in distance from metal ion position to enzyme active site, but it is considered as a polymorphism
-
V9A
-
naturally occuring mutation, the mutant shows 73% reduced activity compared to wild-type, resistant strain
-
D126N
-
site-directed mutagenesis, the mutation causes pyrazinamide resistance, the mutation is located outside of active site and has an allosteric affect
-
D12A
D12G
D49N
E144K
-
naturally occuring mutation from PZA-resistant isolate, analysis of the resistance mechanism of the mutant strain
-
G78C
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
-
K96R
-
naturally occurring mutation in the PncA catalytic region, binding cavity analysis shows an increase of 762.3 A3 in the volume of the mutant protein. Docking studies reveal that pyrazinamide (PZA) has a greater binding affinity for the wild-type protein in comparison to the mutant protein. The residues of flap region acquire more flexibility in mutant form of protein and thus move away from the active site. This leads to weak binding of the drug to the target residues. The mutation leads to a substantial increase in the binding cavity. This prohibits the enzyme from holding the drug properly and therefore pyrazinoic acid (PZA) cannot take its active form
-
N11K
-
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
-
P69T
-
site-directed mutagenesis, the active site mutation causes pyrazinamide resistance, destabilization of the Fe2+ binding site
-
T92C
-
the naturally occuring mutation causes an increase in distance from metal ion position to enzyme active site, but it is considered as a polymorphism
-
V9A
-
naturally occuring mutation, the mutant shows 73% reduced activity compared to wild-type, resistant strain
-
additional information
D12A
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
D12G
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
D49N
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
G78C
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
H51R
naturally occuring mutation, inactive mutant, resistant strain
H51R
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
H57D
naturally occuring mutation, inactive mutant, resistant strain
T135P
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
T142R
naturally occuring mutation, inactive mutant, resistant strain
D12A
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
-
D12G
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
-
D49N
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
-
D12A
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
-
D12G
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type
-
D49N
-
site-directed mutagenesis, the mutation highly reduces the mutant activity compared to wild-type and has a deleterious effect on the metal binding mechanism of PZase
-
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
additional information
analysis of the specific pyrazinamidase (PZase) activity of PncA mutants via PCR-based in vitro-synthesized-PZase assay and correlation of the results to the pyrazinamide (PZA) susceptibility phenotype determined by culture in acidic agar medium at pH 6.0. A set of 23 clinical isolates displaying mutated pncA genes (11 PZA-resistant and 12 PZA-susceptible) and 55 PZA-susceptible clinical strains displaying a wild-type pncA gene are tested. Among the 23 mutants tested, 4 corresponded to mutations not reported before (I5T, Y99S, T142R, and P77L/V131G). Of the 11 PncA mutants expressed from PZA-resistant clinical isolates, 9 are expressed in vitro at yields above 50% relative to the wild-type enzyme. Among them, 6 enzymes (T47P, H51P, H51R, H57D, L85R and T142R) show no detectable activity, while the relative activities for the 3 others, V9A, G97D, and A146V, are low compared to the wild-type PZase. The remaining two mutants, I5T and V9G, presented very low relative expression (5%) and relative activities values of 12 and 1%, respectively. Twelve mutants are expressed from PZA-susceptible isolates. Their expression arte is similar to the wild-type enzyme, and they behave as active pyrazinamidase with specific relative activities ranging from 34 to 314%. Finally, discrepant results are observed for two mutants, V7A and P62T
additional information
-
analysis of the specific pyrazinamidase (PZase) activity of PncA mutants via PCR-based in vitro-synthesized-PZase assay and correlation of the results to the pyrazinamide (PZA) susceptibility phenotype determined by culture in acidic agar medium at pH 6.0. A set of 23 clinical isolates displaying mutated pncA genes (11 PZA-resistant and 12 PZA-susceptible) and 55 PZA-susceptible clinical strains displaying a wild-type pncA gene are tested. Among the 23 mutants tested, 4 corresponded to mutations not reported before (I5T, Y99S, T142R, and P77L/V131G). Of the 11 PncA mutants expressed from PZA-resistant clinical isolates, 9 are expressed in vitro at yields above 50% relative to the wild-type enzyme. Among them, 6 enzymes (T47P, H51P, H51R, H57D, L85R and T142R) show no detectable activity, while the relative activities for the 3 others, V9A, G97D, and A146V, are low compared to the wild-type PZase. The remaining two mutants, I5T and V9G, presented very low relative expression (5%) and relative activities values of 12 and 1%, respectively. Twelve mutants are expressed from PZA-susceptible isolates. Their expression arte is similar to the wild-type enzyme, and they behave as active pyrazinamidase with specific relative activities ranging from 34 to 314%. Finally, discrepant results are observed for two mutants, V7A and P62T
additional information
cumulative effect of mutations and iron substitution
additional information
-
cumulative effect of mutations and iron substitution
additional information
molecular dynamics simulations coupled with essential dynamics and binding pocket analysis at neutral (pH = 7) and acidic (pH = 4) ambient conditions, detailed overview. Computational insights into pH-dependence of structure and dynamics of pyrazinamidase, comparison of wild-type and mutant enzymes
additional information
-
molecular modeling and substrate docking reveals that the wild-type has much stronger binding affinity to PZA than the mutants whereas mutant L151S has the weakest binding affinity. Molecular dynamics simulations and the essential dynamics results illustrate that the positions of the 51st to 71st residues are more dynamics in mutant L151S as compared to the other atoms in PZase. Kinetics of activation and half-life of wild-type and mutant PZases
additional information
-
mutational statistics, especially mutations in the promoter region of pncA, Analysis of methods of phenotypic determination and genotype-phenotype evaluation for diagnostics in clinical drug studies and MIC determinations, overview. For diagnostics, individual mutations (or any subset) are not sufficiently sensitive. Assuming similar error profiles of the 5 phenotyping platforms included in the study, the entire enzyme and its promoter provide a combined estimated sensitivity of 83%. Molecular diagnostics of PZA resistance
additional information
-
cumulative effect of mutations and iron substitution
-
additional information
-
analysis of the specific pyrazinamidase (PZase) activity of PncA mutants via PCR-based in vitro-synthesized-PZase assay and correlation of the results to the pyrazinamide (PZA) susceptibility phenotype determined by culture in acidic agar medium at pH 6.0. A set of 23 clinical isolates displaying mutated pncA genes (11 PZA-resistant and 12 PZA-susceptible) and 55 PZA-susceptible clinical strains displaying a wild-type pncA gene are tested. Among the 23 mutants tested, 4 corresponded to mutations not reported before (I5T, Y99S, T142R, and P77L/V131G). Of the 11 PncA mutants expressed from PZA-resistant clinical isolates, 9 are expressed in vitro at yields above 50% relative to the wild-type enzyme. Among them, 6 enzymes (T47P, H51P, H51R, H57D, L85R and T142R) show no detectable activity, while the relative activities for the 3 others, V9A, G97D, and A146V, are low compared to the wild-type PZase. The remaining two mutants, I5T and V9G, presented very low relative expression (5%) and relative activities values of 12 and 1%, respectively. Twelve mutants are expressed from PZA-susceptible isolates. Their expression arte is similar to the wild-type enzyme, and they behave as active pyrazinamidase with specific relative activities ranging from 34 to 314%. Finally, discrepant results are observed for two mutants, V7A and P62T
-
additional information
-
molecular dynamics simulations coupled with essential dynamics and binding pocket analysis at neutral (pH = 7) and acidic (pH = 4) ambient conditions, detailed overview. Computational insights into pH-dependence of structure and dynamics of pyrazinamidase, comparison of wild-type and mutant enzymes
-
additional information
-
cumulative effect of mutations and iron substitution
-
additional information
-
analysis of the specific pyrazinamidase (PZase) activity of PncA mutants via PCR-based in vitro-synthesized-PZase assay and correlation of the results to the pyrazinamide (PZA) susceptibility phenotype determined by culture in acidic agar medium at pH 6.0. A set of 23 clinical isolates displaying mutated pncA genes (11 PZA-resistant and 12 PZA-susceptible) and 55 PZA-susceptible clinical strains displaying a wild-type pncA gene are tested. Among the 23 mutants tested, 4 corresponded to mutations not reported before (I5T, Y99S, T142R, and P77L/V131G). Of the 11 PncA mutants expressed from PZA-resistant clinical isolates, 9 are expressed in vitro at yields above 50% relative to the wild-type enzyme. Among them, 6 enzymes (T47P, H51P, H51R, H57D, L85R and T142R) show no detectable activity, while the relative activities for the 3 others, V9A, G97D, and A146V, are low compared to the wild-type PZase. The remaining two mutants, I5T and V9G, presented very low relative expression (5%) and relative activities values of 12 and 1%, respectively. Twelve mutants are expressed from PZA-susceptible isolates. Their expression arte is similar to the wild-type enzyme, and they behave as active pyrazinamidase with specific relative activities ranging from 34 to 314%. Finally, discrepant results are observed for two mutants, V7A and P62T
-
additional information
-
molecular dynamics simulations coupled with essential dynamics and binding pocket analysis at neutral (pH = 7) and acidic (pH = 4) ambient conditions, detailed overview. Computational insights into pH-dependence of structure and dynamics of pyrazinamidase, comparison of wild-type and mutant enzymes
-
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
-
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
-
additional information
in silico structural characterizations of pyrazinamidase variants from various species of Mycobacterium
