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
-
-
-
-
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
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
strain Ps49, neomycin phosphotransferase II
-
-
Manually annotated by BRENDA team
-
-
-
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
-
-
-
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+
-
activation at 1 mM, can replace Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
Adenylyl imidodiphosphate
-
-
Gentamicin 1a
kanamycin
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substrate inhibition at 2 mM
tobramycin
additional information
-
not inhibitory: guanylyl imidodiphosphate
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.32
3'-phosphokanamycin
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pH 7.5, 37C, isoenzyme APH3'-IIIa
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0.022
ADP
-
pH 7.5, 37C, isoenzyme APH3'-IIIa
0.0106 - 0.028
ATP
0.3
kanamycin
-
pH 7.5, 35C
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
Escherichia coli
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pH 7.5, 37C, isoenzyme APH3'-IIIa
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
150
ATP
Enterococcus casseliflavus
-
pH 7.5, 37C
4
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0142
Adenylyl imidodiphosphate
-
pH 7.5, 37C
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
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
-
strain ML1629, 5 min, inactivation
55
-
strain H-9, 20 min stable
65
-
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
-
dithiothreitol stabilizes and restores activity of denatured enzyme
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
ammonium sulfate, Sephadex G 50, Sephadex G 100, partial purification
-
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+
-
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+
-
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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