Information on EC 3.1.4.52 - cyclic-guanylate-specific phosphodiesterase

Word Map on EC 3.1.4.52
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Select one or more organisms in this record:
Show additional data
Do not include text mining results
Include (text mining) results (more...)
Include results (AMENDA + additional results, but less precise; more...)

The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
3.1.4.52
-
RECOMMENDED NAME
GeneOntology No.
cyclic-guanylate-specific phosphodiesterase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
cyclic di-3',5'-guanylate + H2O = 5'-phosphoguanylyl(3'->5')guanosine
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of phosphoric diester
SYSTEMATIC NAME
IUBMB Comments
cyclic bis(3->5')diguanylate 3-guanylylhydrolase
Requires Mg2+ or Mn2+ for activity and is inhibited by Ca2+ and Zn2+. Contains a heme unit. This enzyme linearizes cyclic di-3,5-guanylate, the product of EC 2.7.7.65, diguanylate cyclase and an allosteric activator of EC 2.4.1.12, cellulose synthase (UDP-forming), rendering it inactive [1]. It is the balance between these two enzymes that determines the cellular level of c-di-GMP [1].
CAS REGISTRY NUMBER
COMMENTARY hide
338732-46-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
gene binA
-
-
Manually annotated by BRENDA team
gene pdeB
-
-
Manually annotated by BRENDA team
genes bpdA and bpdB
-
-
Manually annotated by BRENDA team
strain KHW
-
-
Manually annotated by BRENDA team
strain KHW
-
-
Manually annotated by BRENDA team
protein CC3396 containing both an EAL and GGDEF motif
-
-
Manually annotated by BRENDA team
gene pdeH
-
-
Manually annotated by BRENDA team
isoform A1
-
-
Manually annotated by BRENDA team
genes 1357c and Rv 1354c
-
-
Manually annotated by BRENDA team
gene perma_0986
-
-
Manually annotated by BRENDA team
gene perma_0986
-
-
Manually annotated by BRENDA team
strain Pf0-1, conditional expression of phosphodiesterase RapA upon exposure to phosphate. Expression is mediated by the Pho regulon
-
-
Manually annotated by BRENDA team
strain Pf0-1, conditional expression of phosphodiesterase RapA upon exposure to phosphate. Expression is mediated by the Pho regulon
-
-
Manually annotated by BRENDA team
PNNL strain, gene pdeB
UniProt
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain El Tor C6706
-
-
Manually annotated by BRENDA team
isoform ScrG, contains GGDEF and EAL domains
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
physiological function
additional information
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
5'-phosphoguanylyl(3'-5')guanosine + H2O
guanosine 5'-monophosphate + guanosine
show the reaction diagram
-
-
-
-
?
5'-phosphoguanylyl(3'->5')guanosine + H2O
GMP + guanosine
show the reaction diagram
the isozyme hydrolyses c-di-GMP in a two-step reaction via the linear intermediate 5'-phosphoguanylyl(3'->5')guanosine, it produces GMP in vitro at a low rate, reaction of EC 3.1.4.1
-
-
?
c-di-GMP + H2O
?
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'-phosphoguanylyl(3'-5')guanosine
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'-phosphoguanylyl(3'->5')guanosine
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'phosphoguanylyl(3'-5')guanosine
show the reaction diagram
cyclic-di-GMP + H2O
5'-pGpG
show the reaction diagram
-
-
-
?
additional information
?
-
NATURAL SUBSTRATES
NATURAL PRODUCTS
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
REVERSIBILITY
r=reversible
ir=irreversible
?=not specified
c-di-GMP + H2O
?
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'-phosphoguanylyl(3'-5')guanosine
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'-phosphoguanylyl(3'->5')guanosine
show the reaction diagram
cyclic di-3',5'-guanylate + H2O
5'phosphoguanylyl(3'-5')guanosine
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Calmodulin
-
-
heme
-
enzyme is regulated via reversible binding of O2 to the heme. Apo-enzyme has less than 2% of phosphodiesterase activity of holo-enzyme. The rate of enzyme autoxidation is the slowest observed so far for this type of heme protein with half-life above 12 h
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
10% of the activity with Mg2+, Mn2+ in presence of Co2+
Fe2+
-
trinuclear Fe binding site located at the bottom of the HD-GYP domain cavity formed by the two claws of the open chela. Residues E185, H189, H221, D222, H250, H276, H277, and D305 are involved in metal binding
Iron
-
heme protein with remarkable stability against electron transfer to O2. Oxy-enzyme is only 30% oxidized to the ferric form after 8 h in airsaturated Tris buffer pH 8.0, 23C
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
5'-phosphoguanylyl-(3'-5')-guanosine
-
the cyclic di-GMP phosphodiesterase activity of YfgFGE and YfgFE is inhibited by the reaction product
Cu2+
-
complete inhibition at 10 mM
GTP
-
inhibition of the phosphodiesterase, reduction of tobramycin biofilm stimulation, and suppression of biofilm response, overview
Nalidixic acid
-
reduction of tobramycin biofilm stimulation and suppression of biofilm response, overview
additional information
-
inhibition of the enzyme leads to suppression of induction of biofilm formation
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
KCl
-
highest catalytic activity in presence of 25 mM KCl
additional information
-
cyclic-di-GMP phosphodiesterase activity of CdpA is regulated by its inactive degenerate GGDEF domain
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.017 - 0.03
5'-phosphoguanylyl(3'->5')guanosine
0.0000029 - 0.119
cyclic di-3',5'-guanylate
0.0003 - 0.0086
cyclic-di-GMP
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00012 - 0.00077
5'-phosphoguanylyl(3'->5')guanosine
0.000011 - 6.7
cyclic di-3',5'-guanylate
0.000011 - 0.67
cyclic-di-GMP
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2.1 - 650
cyclic di-3',5'-guanylate
1294
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.12
-
hydrolysis of 5-phosphoguanylyl(3-5)guanosine, recombinant enzyme, pH 8.0, 30C
2.42
-
synthesis of 5-phosphoguanylyl(3-5)guanosine, recombinant enzyme, pH 8.0, 30C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5 - 9
-
no significant changes in reaction rates within this range
6.2 - 9.5
pH profiles of recombinant wild-type and mutant E352Q enzymes
10
-
slight decrease in activity when raising pH value from 9.4 to 10
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23 - 30
-
-
25
-
assay at
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
additional information
-
anaerobic expression of yfgF is greatest in stationary phase, and in cultures grown at 28C
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
restriction of protein to a single pole requires intact GGDEF and EAL domains
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Caulobacter crescentus (strain NA1000 / CB15N)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
23000
-
2 * 74500 + 2 * 23000, about, sedimentation equilibrium analysis, the dual-functioning diguanylate cyclase/phosphodiesterase enzyme, formed by heme-nitric oxide/oxygen binding protein and cyclic-di-GMP processing enzyme, builds a tetrameric structure
42837
4 * 42837, calculated from amino acid sequence, RocR is mainly a tetramer in solution
47000
-
1 * 47000, calculated, 1 * 47000, sedimentation velocity study
48000
-
gel filtration
74500
-
2 * 74500 + 2 * 23000, about, sedimentation equilibrium analysis, the dual-functioning diguanylate cyclase/phosphodiesterase enzyme, formed by heme-nitric oxide/oxygen binding protein and cyclic-di-GMP processing enzyme, builds a tetrameric structure; 2 * 74500, in solution, sedimentation equilibrium analysis
78000
-
x * 78000, calculated
144300
-
dimeric enzyme, sedimentation equilibrium analysis
176400
calculated from amino acid sequence
197200
-
tetrameric heterotetramer of dual-functioning diguanylate cyclase/phosphodiesterase enzyme complex, sedimentation equilibrium analysis
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
tetramer
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
the REC domain of PA4781 cannot be allosterically activated by phosphorylation in the Escherichia coli background, the REC domain at the N-terminus of the protein is responsible for the phosphodiesterase activity of the enzyme. Phosphorylated and non-phosphorylated proteins are both dimeric
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals are grown by hanging-drop method at 18C using 0.001 ml of protein solution mixed with an equal volume of reservoir solution, varying conditions using 0.1 M bis-Tris, pH 5.5, 0.2-0.4 M ammonium acetate, 25% w/v PEG 3350, with or without 20% v/v DMSO or 2.1 M DL-malic acid, pH 7.0, X-ray diffraction structure determination and analysis at 2.5 A resolution
enzyme does not undergo global changes upon exposure to light. The chromophore environment of full-length enzyme is asymmetric
-
BlrP1 is crystallized at 4C in the dark using polyethyleneglycol in the presence of cyclic-di-GMP and Ca2+
-
purified recombinant detagged enzyme in complex with Fe2+, substrate cyclic di-3',5'-guanylate or final product GMP, hanging drop vapour diffusion method, mixing of equal volumes of 10 mg/ml protein with a precipitant solution made up of 0.1 M MES pH 6.5, 0.9 M succinic acid, and 2% PEG 2000, anomalous X-ray diffraction structure determination and analysis at 2.03-2.8 A resolution
-
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
52
-
loss of activity above
65
-
no residual activity
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
GGDEF domain and GGDEF-EAL fusion domains: GST-affinity chromatography
Ni2+-NTA resin column chromatography; recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant C- and N-terminally His-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration; recombinant wild-type and mutant C- and N-terminally His-tagged enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
recombinant GST-tagged enzyme from Escherichia coli by glutathione affinity chromatography
-
recombinant His-tagged enzyme from Escherichia coli strain BL21(DE3)pLysS cell-free supernatant by nickel affinity chromatography
-
recombinant His-tagged MtbDGC and MtbPDE by nickel affinity chromatography from inclusion bodies
-
recombinant His-tagged protein
-
recombinant N-terminally His6-tagged enzyme from Escherichiia coli strain BL21(DE3) by nickel affinity chromatography and cleavage of the the His-tag by HRV 3C protease
-
recombinant N-terminally maltose-binding protein tagged and C-terminally His6 tagged truncated wild-type enzyme and mutant E364A from Escherichia coli strain Tuner(DE3)pLysS by nicel affinity and amylose affinity chromatography
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
autoinducer-deficient Vibrio cholerae strain transformed with library of random Vibrio cholerae genomic fragments fused to a promoterless luciferase cassette. Verification of Quorum sensing (QS) regulation: wild-type Vibrio cholerae El and QS mutants are transformed with each candidate reporter plasmid, each candidate exhibits similar expression pattern in a luxO D47E strain (response regulator luxO) and a hapR strain (QS transcription factor HapR) that both simulate low cell density, indicating expression of each target gene is dependent on LuxO-P as well as HapR. VCA0681 is overexpressed in Vibrio cholerae luxO D47Eel: reduction of reporter expression. Deletion of either VCA0681 from the Vibrio cholerae El genome does not alter biofilm formation (transcriptional activator vps expression). Absolute level of vps expression in the (HapR minus) Vibrio cholerae El strain 100 times higher than that of wild-type (HapR minus) Vibrio cholerae C1 strain; VCA0895 is overexpressed in Vibrio cholerae luxO D47Eel: growth inhibition, deletion of vca0895 from the Vibrio cholerae El genome does not alter biofilm formation (transcriptional activator vps expression)
-
commercial synthetic gene PA4108, DNA and amino acid sequence determination and analysis, recombinant overexpression of C- and N-terminally His-tagged enzyme in an endogenous PDE yhjH-lacking Escherichia coli strain and in strain BL21(DE3); commercial synthetic gene PA4781, DNA and amino acid sequence determination and analysis, recombinant overexpression of wild-type and mutant C- and N-terminally His-tagged enzymes in an endogenous PDE yhjH-lacking Escherichia coli strain and in strain BL21(DE3), the REC domain of PA4781 cannot be allosterically activated by phosphorylation in the Escherichia coli background
DNA fragments encoding GGDEF (1-172 aa) and GGDEF-EAL domain (10-426 aa) amplified separately and fused to the GST coding region in pGEX-6p-1, transformation of Escherichia coli BL21. Deletion of Xcc1959 in Xcc strain XC1 or 8004 had no significant effect on virulence factor production in vitro; mutated DAA domain cloned in expression vector pLAFR3 (lac promoter) for complementation analysis in a Xanthomonas campestris XC1 mutant lacking ravR. Deletion of ravR results in decreased virulence factor production. RavS and RavR constitute a two-component regulatory system (ravS gene deleted inframe in Xanthomonas campestris strains XC1 and 8004): no effect on bacterial growth in YEB and LB medium, similar phenotype changes compared to Xanthomonas campestris XC1 mutant lacking ravR. RavR is a c-di-GMP phosphodiesterase: fusionated domains GGDEF and EAL (cloned separately into the expression vector pLAFR3 for in trans expression in the mutant deltaravR). RavS/RavR 2-component system regulates diverse functions in Xanthomonas campestris XC1
ectopic expression of 31 of the conserved genes from 20 genomes of Clostridium difficile for comparison of their effect on motility and biofilm formation. Most of the PDEs are active in a Vibrio cholerae model
-
expressed in Escherichia coli strain BL21; gene PA3947, expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression in Escherichia coli with His-tag
-
expression in Escherichia coli, His-tag
-
Flagellar promoter analysis in the bpdA, bpdB, and cgsB mutants using a luminescent reporter system. Expression in Vibrio parahaemolyticus strain LM5984
-
gene pdeB, gene expression profiling and quantitative PCR in wild-type and mutant cells, expression of GFP-tagged mutant enzymes, expression of N-terminally maltose-binding protein tagged and C-terminally His6 tagged truncated wild-type enzyme and mutant E364A in Escherichia coli strain Tuner(DE3)pLysS
gene perma_0986, expression of N-terminally His6-tagged enzyme in Escherichiia coli strain BL21 (DE3)
-
gene rbdA, expression as GST-tagged enzyme in Escherichia coli
-
gene yfgF is expressed under anaerobic conditions from a class II FNR, i.e. regulator of fumarate and nitrate reduction-dependent promoter
-
genes 1357c and Rv 1354c, expression of enzymes MtbDGC and MtbPDE as His-tagged proteins in inculsion bodies
-
overexpression in Escherichia coli
-
overexpression is sufficient to induce lateral flagellar gene expression in liquid, decrease biofilm formation, decrease cps gene expression, and suppress the DeltascrABC phenotype
-
overexpression of BinA in Vibrio fischeri enhancing motility. Expression of wild-type and mutant enzymes in Escherichia coli
-
overexpression with His-tag
-
recombinant expression of genetic constructs in Bacillus subtilis strain DS2569 to generate phage lysates for transduction
-
recombinant expression of His-tagged enzyme in Escherichia coli strain BL21(DE3)pLysS
-
sequence comparisons, comparative transcriptional profiling of Bd1817 expression
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
cdpA is expressed during in vitro growth in a biofilm but is not expressed in vivo until the late stage of infection, after colonization has occurred
-
cdpA is expressed during in vitro growth in a biofilm but is not expressed in vivo until the late stage of infection, after colonization has occurred, cdpA expression is not regulated by cyclic-di-GMP concentration
-
low growth rates promote yfgF expression
-
overexpression of the syp regulator sypG induces binA expression
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
E323Q
-
mutation in EAL motif, both catalytic activity and induction by GTP are not affected
L128C/G353C
-
the mutant shows higher catalytic efficiency at pH 7.5 and 20C compared to the wild type enzyme under light and dark conditions
R93S
-
the mutant shows higher catalytic efficiency at pH 7.5 and 20C compared to the wild type enzyme under light and dark conditions
S309C/S312C
-
inactive at pH 7.5 and 20C under dark and light conditions
C406S
-
site-directed mutagenesis of enzyme MtbDGC, the mutant is catalytically inactive
D183A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
D222A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
D305A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
E185A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
G284A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
H189A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
H221A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
H250A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
H276A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
H277A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
K225A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
K317A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
P286A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
R314 A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
Y285 A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
D305A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
-
G284A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
-
H189A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
-
H221A
-
site-directed mutagenesis, the mutant shows highly reduced catalytic activity compared to the wild-type enzyme
-
P286A
-
site-directed mutagenesis, the mutant shows reduced catalytic activity compared to the wild-type enzyme
-
D295A
inactive; site-directed mutagenesis, inactive mutant
D318A
site-directed mutagenesis, the mutant shows a reduced kcat and altered Km compared to the wild-type enzyme; the mutant shows 8.4fold reduction in turnover number
D347A
-
mutant is indistinguishable from enzyme null mutant. Loss of twitching activity and PO4 phage sensitivity, mutant fails to assemble surface pili
D56N
-
the RocR mutant exhibits a significantly smaller Km value than that of the wild type enzyme
E175A
inactive; site-directed mutagenesis, inactive mutant
E265A
inactive; site-directed mutagenesis, inactive mutant
E314A
site-directed mutagenesis
E352A
inactive; site-directed mutagenesis, inactive mutant
E352C
inactive; site-directed mutagenesis, inactive mutant
E352D
site-directed mutagenesis, the mutant shows a reduced kcat and altered Km compared to the wild-type enzyme; the mutant shows 30000fold reduction in turnover number
E352Q
site-directed mutagenesis, the mutant shows a reduced kcat and altered Km compared to the wild-type enzyme; the mutant shows 61000fold reduction in turnover number
E355A
site-directed mutagenesis, the mutant shows a 1.3fold reduced kcat and increased Km compared to the wild-type enzyme; the mutant shows 1.3fold reduction in turnover number
E464A
-
the PA2567 mutant shows increased kcat and Km compared to the wild type enzyme
E475A
-
mutant is indistinguishable from enzyme null mutant. Loss of twitching activity and PO4 phage sensitivity, mutant fails to assemble surface pili
F297A
-
the RocR mutant shows decreased kcat and Km compared to the wild type RocR enzyme
F498A
-
the PA2567 mutation reduces the enzyme activity to below the measurable level
G346A
-
mutant is indistinguishable from enzyme null mutant. Loss of twitching activity and PO4 phage sensitivity, mutant fails to assemble surface pili
K316A
inactive; site-directed mutagenesis, inactive mutant
L477A
-
mutant is indistinguishable from enzyme null mutant. Loss of twitching activity and PO4 phage sensitivity, mutant fails to assemble surface pili
N233A
inactive; site-directed mutagenesis, inactive mutant
Q161A
site-directed mutagenesis, the mutant shows increased Km compared to the wild-type enzyme; the mutant shows 5.1fold reduction in turnover number
Q372A
site-directed mutagenesis, the mutant shows a reduced kcat and altered Km compared to the wild-type enzyme; the mutant shows 8.4fold reduction in turnover number
R179A
site-directed mutagenesis, the mutant shows a 29.1fold reduced kcat and increased Km compared to the wild-type enzyme; the mutant shows 29.1fold reduction in turnover number
S302A
-
the RocR mutant shows decreased kcat and increased Km compared to the wild type RocR enzyme
S493A
-
the PA2567 mutation reduces the enzyme activity to below the measurable level
T267A
site-directed mutagenesis, the mutant shows a reduced kcat and altered Km compared to the wild-type enzyme; the mutant shows 13.4fold reduction in turnover number
V476A
-
mutant is indistinguishable from enzyme null mutant. Loss of twitching activity and PO4 phage sensitivity, mutant fails to assemble surface pili
E634A
site-directed mutagenesis
D289A
-
VCA0681, mutation in active site, eliminated VCA0681 repression of vpsL-lux, biofilm formation not repressed by mutant contrary to wild-type. Expression of the active-site mutant (D289A) in a luxO D47E strain has little effect on cyclic di-3',5'-guanylate levels
E170A
-
mutation in metal binding site, loss of activity
D289A
-
VCA0681, mutation in active site, eliminated VCA0681 repression of vpsL-lux, biofilm formation not repressed by mutant contrary to wild-type. Expression of the active-site mutant (D289A) in a luxO D47E strain has little effect on cyclic di-3',5'-guanylate levels
-
D224A
-
mutation in HDDDF motif, decrease in activity
D225A
-
mutation in HDDDF motif, strong decrease in activity
E350A
-
mutation in EAL motif, almost complete loss of activity
F226A
-
mutation in HDDDF motif, activity similar to wild-type
additional information
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
refolding of recombinant His-tagged MtbDGC and MtbPDE after purification from inclusion bodies via stepwise dialysis
-
Show AA Sequence (5011 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.