Information on EC 2.7.1.95 - kanamycin kinase

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The expected taxonomic range for this enzyme is: Bacteria, Eukaryota

EC NUMBER
COMMENTARY hide
2.7.1.95
-
RECOMMENDED NAME
GeneOntology No.
kanamycin kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + kanamycin = ADP + kanamycin 3'-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
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-
-
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SYSTEMATIC NAME
IUBMB Comments
ATP:kanamycin 3'-O-phosphotransferase
Also acts on the antibiotics neomycin, paromomycin, neamine, paromamine, vistamycin and gentamicin A. An enzyme from Pseudomonas aeruginosa also acts on butirosin.
CAS REGISTRY NUMBER
COMMENTARY hide
62213-36-9
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
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-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
neomycin phosphotransferase II
-
-
Manually annotated by BRENDA team
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + kanamycin
ADP + kanamycin 3'-phosphate
show the reaction diagram
ATP + neamine
ADP + neamine phosphate
show the reaction diagram
-
-
-
?
ATP + neomycin
ADP + neomycin 3'-phosphate
show the reaction diagram
ATP + neomycin
ADP + neomyin phosphate
show the reaction diagram
ATP + paromomycin
ADP + paromomycin phosphate
show the reaction diagram
-
-
-
?
additional information
?
-
-
under physiologic concentrationsof NTPs, enzyme exclusively uses ATP
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-
-
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 + neomycin
ADP + neomycin 3'-phosphate
show the reaction diagram
-
-
-
-
?
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
activation at 1 mM, can replace Mg2+
Mn2+
-
activation at 1 mM, can replace Mg2+
Zn2+
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activation at 1 mM, can replace Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Adenylyl imidodiphosphate
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Gentamicin 1a
kanamycin
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substrate inhibition at 2 mM
tobramycin
additional information
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not inhibitory: guanylyl imidodiphosphate
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KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.32
3'-phosphokanamycin
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pH 7.5, 37°C, isoenzyme APH3'-IIIa
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0.022
ADP
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pH 7.5, 37°C, isoenzyme APH3'-IIIa
0.0106 - 0.028
ATP
0.3
kanamycin
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pH 7.5, 35°C
0.004 - 0.013
kanamycin A
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.11 - 1.76
ATP
1.79
kanamycin A
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pH 7.5, 37°C, isoenzyme APH3'-IIIa
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
150
ATP
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pH 7.5, 37°C
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0142
Adenylyl imidodiphosphate
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pH 7.5, 37°C
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
7.5
-
with kanamycin
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 8
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approx. half-maximal activity at pH 6.0 and pH 8.0
6.5 - 8
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approx. half-maximal activity at pH 6.5 and pH 8.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
35 - 37
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
transformed with pAG60
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
PDB
SCOP
CATH
UNIPROT
ORGANISM
Acinetobacter baumannii (strain AYE);
Enterococcus faecalis;
P00552
Klebsiella pneumoniae;
Q9F9M5
Streptomyces rimosus;
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phosphoprotein
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
7.5 - 9
-
stable
645102
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
45
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strain ML1629, 5 min, inactivation
55
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strain H-9, 20 min stable
65
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strain H-9, 5 min stable
75
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strain H-9, inactivation
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
ammonium chloride or neomycin B stabilizes, E. coli JR39 enzyme
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dithiothreitol stabilizes and restores activity of denatured enzyme
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, Sephadex G 50, Sephadex G 100, partial purification
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expressed in COS-1 cells
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expression of kanaR gene in Escherichia coli
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expression of neomycin phosphotransferase II in Nicotiana tobacco
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expression of neomycin phosphotransferase II-glycophorin A fusion protein in Escherichia coli and CHO cells
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expression of neoR gene in Trypanosoma cruzi
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overexpression of aminoglycoside phosphotransferase-IIIa, i.e. APH3'-IIIa in Escherichia coli
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
S146A
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19% of the activity of phosphorylated wild-type, 90% of the activity of unphosphorylated wild-type, in presence of Ca2+
S146A/S160A
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17% of the activity of phosphorylated wild-type, 85% of the activity of unphosphorylated wild-type, in presence of Ca2+
S146A/S160A/S215A
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17% of the activity of phosphorylated wild-type, 60% of the activity of unphosphorylated wild-type, in presence of Ca2+
S215A
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88% of the activity of phosphorylated wild-type, 67% of the activity of unphosphorylated wild-type, in presence of Ca2+
S95A/S160A
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82% of the activity of phosphorylated wild-type, 95% of the activity of unphosphorylated wild-type, in presence of Ca2+
S146A
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19% of the activity of phosphorylated wild-type, 90% of the activity of unphosphorylated wild-type, in presence of Ca2+
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S146A/S160A
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17% of the activity of phosphorylated wild-type, 85% of the activity of unphosphorylated wild-type, in presence of Ca2+
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S146A/S160A/S215A
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17% of the activity of phosphorylated wild-type, 60% of the activity of unphosphorylated wild-type, in presence of Ca2+
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S215A
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88% of the activity of phosphorylated wild-type, 67% of the activity of unphosphorylated wild-type, in presence of Ca2+
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S95A/S160A
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82% of the activity of phosphorylated wild-type, 95% of the activity of unphosphorylated wild-type, in presence of Ca2+
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APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
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competitive assay that mimics in vivo nucleotide triphosphate concentrations and usage by the enzyme. Downstream analysis of reaction products by high-performance liquid chromatography enables the determination of partitioning of phosphate flux from nucleotide triphosphate donors to antibiotics
medicine
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the enzyme can be used in immunoliposome-mediated delivery for the lineage-specific selection of differentiated/committed stem cell progenies for transplantation, method development and evaluation, potential advantages over transfection with marker genes, fluorescence-activated and magnetic affinity cell-sorting, overview
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