Information on EC 2.7.1.78 - polynucleotide 5'-hydroxyl-kinase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.7.1.78
-
RECOMMENDED NAME
GeneOntology No.
polynucleotide 5'-hydroxyl-kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + 5'-dephospho-DNA = ADP + 5'-phospho-DNA
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phopho group transfer
phospho group transfer
-
-
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:5'-dephosphopolynucleotide 5'-phosphotransferase
Also acts on 5'-dephospho-RNA 3'-mononucleotides.
CAS REGISTRY NUMBER
COMMENTARY hide
37211-65-7
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
from infected Escherichia coli
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
chinese hamster lung cells
-
-
Manually annotated by BRENDA team
expression in Escherichia coli with His10-tag
-
-
Manually annotated by BRENDA team
expression in Escherichia coli with His10-tag
-
-
Manually annotated by BRENDA team
no activity in bacteriophage T1
-
-
-
Manually annotated by BRENDA team
no activity in bacteriophage T5
-
-
-
Manually annotated by BRENDA team
no activity in Escherichia coli
-
-
-
Manually annotated by BRENDA team
infecting Rhodothermus marinus, bifunctional enzyme with 5-kinase domain and 3-phosphohydrolase domain
-
-
Manually annotated by BRENDA team
synthetic ribozyme
class I polynucleotide kinase
-
-
Manually annotated by BRENDA team
trifunctional enzyme with RNA ligase, polynucleotide 5'-kinase, and polynucleotide 3'-phosphatase activities
-
-
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
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
ATP + (AG)10GGCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + (AG)10GGCCC-tetramethylrhodamine
?
show the reaction diagram
-
-
-
?
ATP + 2'(3')-ribonucleotides
?
show the reaction diagram
ATP + 3'-CMP
ADP + pCp
show the reaction diagram
ATP + 5'-dephospho-DNA
ADP + 5'-phospho-DNA
show the reaction diagram
ATP + 5'-dephospho-RNA
ADP + 5'-phospho-RNA
show the reaction diagram
ATP + 5'-dsRNA
ADP + 5'-phospho-dsRNA
show the reaction diagram
-
-
-
-
?
ATP + 5'-OH DNA
ADP + 5'-phospho-DNA
show the reaction diagram
-
-
-
r
ATP + 5'-OH RNA
ADP + 5'-phospho-RNA
show the reaction diagram
preference for RNA substrates
-
-
r
ATP + d(A20)
ADP + 5'-phospho-d(A20)
show the reaction diagram
-
-
-
-
?
ATP + deoxynucleoside 3'-phosphate
ADP + deoxynucleoside 3',5'-diphosphate
show the reaction diagram
-
-
-
-
?
ATP + GGGCC(AG)10GGCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + GGGCC(AG)10GGCCC-tetramethylrhodamine
?
show the reaction diagram
-
-
-
?
ATP + GGGCC(AG)12GGCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + GGGCC(AG)8GGCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + GGGGC(AG)10GCCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + GGGGC(AG)10GCCCC-tetramethylrhodamine
?
show the reaction diagram
-
-
-
?
ATP + GGGGG(AG)10CCCCC-fluorescein
?
show the reaction diagram
-
-
-
?
ATP + GGGGG(AG)10CCCCC-tetramethylrhodamine
?
show the reaction diagram
-
-
-
?
ATP + nucleoside-3'-monophosphate
ADP + nucleoside-3',5'-diphosphate
show the reaction diagram
ATP + r(A20)
ADP + 5'-phospho-r(A20)
show the reaction diagram
-
-
-
-
?
ATP + synthetic oligonucleotide
ADP + oligonucleotide 5'-phosphate
show the reaction diagram
beta,gamma-imidoadenylyl 5'-triphosphate + 5'-phospho-DNA
beta,gamma-imidoadenylyl 5'-tetraphosphate + 5'-dephospho-DNA
show the reaction diagram
CTP + 5'-dephospho-DNA
CDP + 5'-phospho-DNA
show the reaction diagram
CTP + 5'-dephospho-RNA
CDP + 5'-phospho-RNA
show the reaction diagram
-
less effective than ATP
-
?
dATP + 5'-dephospho-DNA
dADP + 5'-phospho-DNA
show the reaction diagram
-
-
-
?
GTP + 5'-dephospho-DNA
GDP + 5'-phospho-DNA
show the reaction diagram
TTP + 5'-dephospho-DNA
TDP + 5'-phospho-DNA
show the reaction diagram
-
-
-
?
UTP + 5'-dephospho-DNA
UDP + 5'-phospho-DNA
show the reaction diagram
[gamma-S]ATP + 5'-dephospho-RNA
[gamma-S]ADP + 5'-phospho-RNA
show the reaction diagram
-
-
-
?
[gamma-S]ATP + 5'-OH-(ribonucleotide)7
[gamma-S]ADP + 5'-phospho-(ribonucleotide)7
show the reaction diagram
[gamma-S]GTP + 5'-dephospho-RNA
[gamma-S]GDP + 5'-phospho-RNA
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
ATP + 5'-dephospho-DNA
ADP + 5'-phospho-DNA
show the reaction diagram
ATP + 5'-dephospho-RNA
ADP + 5'-phospho-RNA
show the reaction diagram
ATP + 5'-dsRNA
ADP + 5'-phospho-dsRNA
show the reaction diagram
-
-
-
-
?
ATP + 5'-OH DNA
ADP + 5'-phospho-DNA
show the reaction diagram
Q57936
-
-
-
r
ATP + 5'-OH RNA
ADP + 5'-phospho-RNA
show the reaction diagram
Q57936
preference for RNA substrates
-
-
r
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
4-(3-bromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-yl)-N1-cyclohexyl-N2-[2-(1H-imidazol-4-yl)ethyl]piperazine-1,2-dicarboxamide
4-(3-bromo-8-chloro-6,11-dihydro-5H-benzo[5,6]cyclohepta[1,2-b]pyridin-11-yl)-N1-cyclohexyl-N2-[3-(1H-imidazol-4-yl)propyl]piperazine-1,2-dicarboxamide
-
-
ADP
-
reverse reaction
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Divalent cations
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
2-(1-hydroxyundecyl)-1-(4-nitrophenylamino)-6-phenyl-6,7a-dihydro-1H-pyrrolo[3,4-b]pyridine-5,7(2H,4aH)-dione
2-(hydroxy(3,4,5-trimethoxyphenyl)methyl)-1-(4-nitrophenylamino)-6-phenyl-6,7a-dihydro-1H-pyrrolo[3,4-b]pyridine-5,7(2H,4aH)-dione
-
A6B4C3
2-(hydroxy(phenyl)methyl)-1-(4-nitrophenylamino)-6-phenyl-6,7a-dihydro-1H-pyrrolo[3,4-b]pyridine-5,7(2H,4aH)-dione
-
A1B4C3
2-(hydroxy(thiophen-2-yl)methyl)-6-methyl-1-(phenylamino)-6,7a-dihydro-1H-pyrrolo[3,4-b]pyridine-5,7(2H,4aH)-dione
-
A39B1C2
5'-AMP
-
weak
5'-Hydroxyl poly(I)
-
in combination with 5'-hydroxyl poly(A) or poly(C)
AgNO3
ammonium sulfate
beta,gamma-imidoadenosine 5'-triphosphate
-
binds with high affinity, similar to ATP
beta,gamma-imidoadenylyl 5'-tetraphosphate
-
ATP analog, competitive against ATP, noncompetitive against 5'-OH-DNA, serves as substrate for the reverse reaction only
Ca2+
-
inhibits in combination with MgCl2, stimulates without MgCl2
Cd2+
-
-
Chloramphenicol
Cibacron blue F3GA
-
chromophore of blue dextran, inhibition is competitive to single stranded DNA, noncompetitive with respect to ATP
CTP
-
more than 90% inhibition at 0.3 mM
dATP
-
more than 95% inhibition at 0.3 mM
dCTP
-
80% inhibition at 0.3 mM
Deoxyribonucleoside triphosphates
-
-
Dextran sulfate
-
dGTP
-
more than 95% inhibition at 0.3 mM
diphosphate
dTTP
-
more than 95% inhibition at 0.3 mM
GTP
-
more than 90% inhibition at 0.3 mM
heparin
iodoacetate
N-ethylmaleimide
-
-
Ni2+
-
inhibits in combination with MgCl2, stimulates without MgCl2
p-chloromercuribenzoate
p-hydroxymercuribenzoate
-
2-mercaptoethanol prevents inhibition
PEG 6000
-
5-15%, 3-4fold increase in activity, inhibitory above
phosphate
Ribonucleoside 3'-phosphates
-
weak
Ribonucleoside triphosphates
-
-
spermine
-
1 mM enhances activity 3times, inhibition above 1 mM
sulfate
sulfhydryl antagonists
-
-
-
tert-butyl 2-(1-hydroxy-2,2-diphenylethyl)-6-methyl-5,7-dioxo-2,4a,5,6,7,7a-hexahydro-1H-pyrrolo[3,4-b]pyridine-1yl-carbamate
-
A26B11C2
UTP
-
more than 90% inhibition at 0.3 mM
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
bovine serum albumin
-
dithiothreitol
PEG 6000
-
5-15%, 3-4fold increase in activity, inhibitory above
Polyamines
-
Polyethylene glycol
-
4-10%, stimulates
spermidine
spermine
sulfhydryl reagents
XRCC1
-
XRCC4
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0025
5'-hydroxyl poly(A)
-
pH 8.4, 37C
0.0034
5'-hydroxyl poly(C)
-
pH 8.4, 37C
0.016
5'-OH DNA
-
0.0027 - 0.046
5'-OH-DNA
0.021
5'-phospho-DNA
-
0.2
ADP
-
dephosphorylation of single-stranded oligonucleotides, pH 7.6, 37C; phosphate exchange reaction between nucleotides, reverse reaction, pH 7.6, 37C
0.00069 - 3.4
ATP
0.00056
GGGCC(AG)10GGCCC-fluorescein
-
-
0.5 - 2.5
Mg2+
0.1
Mn2+
-
pH 6.0, 37C
0.008
oligo (dT)25
-
polymin P precipitable enzyme, pH 7.5, 37C
-
0.0222 - 0.143
oligonucleotides
-
depending on the 5'-base and the length of the oligonucleotide, pH 7.6, 37C
0.0036
Poly(A)
-
pH 8.4, 37C
0.0013 - 0.0015
r(A20)
-
0.22 - 3.1
[gamma-S]ATP
0.53
[gamma-S]GTP
-
pH 8.4, 37C
additional information
additional information
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.008 - 1.6
3'-CMP
0.077
5'-OH DNA
Pyrococcus horikoshii
Q57936
-
0.00083
5'-phospho-DNA
Pyrococcus horikoshii
Q57936
-
0.00005 - 0.2
ATP
198
GGGCC(AG)10GGCCC-fluorescein
Enterobacteria phage T4
P06855
-
-
1.9 - 2.6
r(A20)
-
0.00283
[gamma-S]ATP
synthetic ribozyme
-
pH-independent, 30C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3.8
ADP
synthetic ribozyme
-
pH-independent in the range pH 6.0-8.5, 30C
3 - 11
beta,gamma-imidoadenylyl 5'-tetraphosphate
0.2 - 5
diphosphate
0.5 - 7
sulfate
additional information
additional information
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.00006
2-(1-hydroxyundecyl)-1-(4-nitrophenylamino)-6-phenyl-6,7a-dihydro-1H-pyrrolo[3,4-b]pyridine-5,7(2H,4aH)-dione
Homo sapiens
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0002
-
purified, polymin P precipitable enzyme, pH 5.5, 37C
0.0007
-
partially purified enzyme, substrate RNA
0.0012
-
purified enzyme
0.0028
-
purified enzyme
0.0043
-
purified enzyme
0.005
-
above, purified enzyme, 30C
0.006
-
partially purified enzyme
0.0167
-
purified enzyme
0.0418
-
purified enzyme
0.125
-
purified enzyme, substrate RNA
0.393
-
purified enzyme
0.53
-
purified enzyme
2.67
-
substrate r(A20), pH 6.0, 65C
3.67
-
substrate d(A20), pH 6.0, 65C
5.67
-
purified wild-type
7
-
purified mutant pseT 1
30 - 40
-
commercial preparation
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5
reverse reaction
5.5 - 7.5
-
3-CMP kinase activit
6 - 9
-
broad optimum
6.2
-
imidazole buffer
7 - 9
-
polymin P precipitable enzyme
7.8
-
inhibition assay
7.9 - 8.9
-
-
8.4
-
assay at, substrate RNA
9
-
substrate RNA
additional information
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
50% of maximum activity
4.5 - 6.5
RNA kinase activity
4.5 - 9.5
DNA kinase activity
4.5 - 7
5 - 7
-
pH 5: about 5% of activity maximum, pH 7: about 30% of activity maximum
5 - 9.5
-
about 50% of activity maximum at pH 5.0 and pH 9.5
5 - 6.5
-
pH 5: about 50% of activity maximum, pH 6.5: about 35% of activity maximum
6.5 - 8.9
-
pH 6.5: about 50% of activity maximum, pH 7.9-8.9: activity maximum, substrate RNA
7.4 - 8
-
forward reaction, optimal range
7.5 - 9.4
-
pH 7.5: about 50% of activity maximum, pH 8.4 and pH 9.4: about 85% of activity maximum
7.5 - 10
-
more than 80% of maximum activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
25 - 30
-
-
additional information
synthetic ribozyme
-
detailed reaction kinetics, pH- and temperature-dependence
TEMPERATURE RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0 - 37
-
0C: about 7% of activity maximum, 37C: activity maximum, bacteriophage T4 enzyme
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
8.5
-
-
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
ORGANISM
UNIPROT
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Clostridium thermocellum (strain ATCC 27405 / DSM 1237 / NBRC 103400 / NCIMB 10682 / NRRL B-4536 / VPI 7372)
Saccharomyces cerevisiae (strain ATCC 204508 / S288c)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
38000
-
gel filtration
41000
determined by SDS-PAGE, tag-free
56000
-
gel filtration
57100
-
-
58000
-
sedimentation equilibrium analysis
70000
-
gel filtration, sucrose density gradient centrifugation
72000
-
sucrose density gradient centrifugation
79000
-
gel filtration
80000 - 160000
80000
-
gel filtration
96000
-
gel filtration, native PAGE
140000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
homodimer
crystal structure
monomer
tetramer
additional information
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
crystals soaked with DNA oligonucleotides
crystal structure determination and analysis
-
inactivated PNK mutant protein with several 3'-phosphorylated DNAs of different sequence bound in the phosphatase active site
-
kinase domain of enzyme Pnkp bound to ATP-Mg2+ or ADP-Mg2+, by hanging drop vapor diffusion, protein solution containing 0.6 mM kinase, 2 mM ATP, and 10 mM MgCl2 with an equal volume of precipitant solution containing 100 mM HEPES, pH 7.5, 200 mM MgCl2, and 30% PEG 400, 22C, 2 days, X-ray diffraction structure determination and analysis at 2.1 A resolution
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5
-
37C, unstable below
642022
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
55
-
5 min, complete loss of activity
70
-
stable for 1 h
additional information
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ATP protects against heat denaturation and tryptic digestion
-
bovine serum albumin and dithiothreitol stabilize the enzyme during activity assay
-
freezing and thawing reduces activity after 1 cycle by 60%
greatest protection from heat inactivation with substrates for the kinase, e.g. thymus DNA and ribosomal RNA, both micrococcal nuclease-treated and 3'-AMP
-
high ionic strength, 0.1 M KCl, spermine, ATP and thymidine 3'-monophosphate stabilize the oligomeric structure
-
high ionic strength, e.g. 0.1 M KCl, spermine, ATP and thymidine 3'-monophosphate are essential to stabilize the enzyme in an oligomeric state
-
protection against heat inactivation by ATP or 5'-OH-DNA at low and moderately high NaCl concentrations
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 10 mM Na2HPO4, pH 7.4, 50% v/v glycerol, 1 mM DTT, 0.001 mM ATP, stable for 2 months
-
-20C, 50 mM Tris-HCl, pH 7.5, 25 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.001 mM ATP, 50% glycerol, stable for up to 18 months
-
0-4C, stable for at least 1 week
-
0C, 10% loss of activity after 2 months
-
0C, partially purified ammonium sulfate fraction, stable for more than 1 year
-
0C, partially purified enzyme, 50 mM Tris-HCl, pH 7.5, 30 mM NaCl, 0.5 mM DTT, 0.5 mM EDTA, on ice, stable for several weeks
-
0C, purified protein, concentration 0.014 mg/ml, stable for 5 months
-
0C, stable for at least 1 month
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
10860fold, to near homogeneity
-
1500fold
-
2 mutant forms pseT 1 and pseT 47
-
431fold
-
6717fold, to near homogeneity
-
Ni-agarose chromatography
-
Ni-NTA agarose column chromatography
-
nickel-agarose chromatography
-
on a Ni-NTA-agarose column, the His10Smt3 tag is removed with the Smt3-specific protease Ulp1
over 1875fold, to homogeneity
-
partial, 35fold
-
partial, 960fold
-
polymin P precipitable enzyme, 1570fold
-
recombinant GST-tagged human Nol9 from Sf9 insect cells by glutathione affinity chromatography
-
recombinant His-tagged full length enzyme and recombinant truncated enzyme from Escherichia coli
-
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3) by nickel affinity chromatography
recombinant N-terminally His-tagged polynucleotide kinase and aprataxin-like forkhead-associated from Escherichia coli strain BL21(DE3) by nickel affinity chromatography and gel filtration
-
the mutant proteins are purified using ProBond resin
to homogeneity
-
to near homogeneity
-
using Strep-Tactin resin
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
a vector encoding N-terminally Strep II epitope-tagged polynucleotide kinase is prepared, the protein is expressed in Escherichia coli Rosetta 2 DE3 pLysS cells
-
expressed in Escherichia coli
-
expressed in Escherichia coli Rosetta2(DE3)pLysS cells
-
expression as soluble His-tagged protein in Escherichia coli; expression of truncated enzyme, residue Met140 to C-terminus, in Escherichia coli
-
expression of GST-tagged human Nol9 in Spodoptera frugiperda Sf9 insect cells
-
expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)
expression of N-terminally His-tagged polynucleotide kinase and aprataxin-like forkhead-associated in Escherichia coli strain BL21(DE3)
-
into the vector pET28-His10Smt3 for expression in Escherichia coli BL21DE3 cells
the PNKP mutants are subcloned into the vector pET16b for expression in Escherichia coli BL21DE3 pLysS cells
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D165A
-
site-directed mutagenesis, increased activity compared to the wild-type, no 3'-phosphatase activity remaining
D180A
-
site-directed mutagenesis, slightly higher activity than the wild-type, reduced 3'-phosphatase activity
D250A
-
site-directed mutagenesis, increased activity compared to the wild-type , reduced 3'-phosphatase activity
D254A
-
site-directed mutagenesis, increased activity compared to the wild-type, nearly no 3'-phosphatase activity remaining
D278A
-
site-directed mutagenesis, increased activity compared to the wild-type, no 3'-phosphatase activity remaining
K125A
-
site-directed mutagenesis, slightly higher activity than the wild-type
R122A
-
site-directed mutagenesis, slightly higher activity than the wild-type, reduced 3'-phosphatase activity
R126A
-
site-directed mutagenesis, highly reduced activity
R176A
-
site-directed mutagenesis, increased activity compared to the wild-type, highly reduced 3'-phosphatase activity
R213A
-
site-directed mutagenesis, increased activity compared to the wild-type, no 3'-phosphatase activity remaining
R246A
-
site-directed mutagenesis, increased activity compared to the wild-type, highly reduced 3'-phosphatase activity
R279A
-
site-directed mutagenesis, increased activity compared to the wild-type, highly reduced 3'-phosphatase activity
R36A
-
site-directed mutagenesis, similar activity than the wild-type
R38A
-
site-directed mutagenesis, highly reduced activity, reduced 3'-phosphatase activity
S84A
-
site-directed mutagenesis, slightly lower activity than the wild-type, reduced 3'-phosphatase activity
D171A
-
complete loss of activity
D173A
-
complete loss of activity
K127A
-
inactive
K378A
-
complete loss of activity
W331F
single mutant, 90% phosphatase activity
W402F
single mutant, 85% phosphatase activity
WFX402
mutant, all tryptophans except 402 are replaced by phenylalanine, 85% phosphatase activity
D169A
-
50% of wild-type activity with 37-mer oligodeoxyribonucleotide substrate, no reverse reaction
K15A
-
no enzymic activity in kinase reaction, 58% of wild-type activity in 3-phosphates reaction
D175A
-
60% of wild-type activity with 37-mer oligodeoxyribonucleotide substrate, 68% of wild-type activity with CMP substrate
K16A
-
no enzymic activity
D73A
mutation abolishes activity
K49A
mutation reduces activity to 1% of the wild-type value
D38E
site-directed mutagenesis, the mutant shows 6% of wild-type activity
D38N
site-directed mutagenesis, inactive mutant
H189D
-
mutation transforms enzyme into a Mn2+-dependent phosphodiesterase devoid of monoesterase activity. Phosphodiesterase activity of mutant is strictly specific for 2',3'-cyclic phosphates which it hydrolyzes to yield a 2'-NMP as the sole product
K21Q
site-directed mutagenesis, the mutant shows 100fold reduced activity compared to the wild-type enzyme
K21R
site-directed mutagenesis, the mutant shows 15fold reduced activity compared to the wild-type enzyme
K95A
site-directed mutagenesis, inactive mutant, the phenotype is benign
R41K
site-directed mutagenesis, the mutant shows about 65% reduced activity compared to the wild-type enzyme
R41Q
site-directed mutagenesis, the mutant shows 100fold reduced activity compared to the wild-type enzyme
S22T
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
additional information