Information on EC 2.7.1.35 - pyridoxal kinase

Word Map on EC 2.7.1.35
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 expected taxonomic range for this enzyme is: Eukaryota, Bacteria

EC NUMBER
COMMENTARY hide
2.7.1.35
-
RECOMMENDED NAME
GeneOntology No.
pyridoxal kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + pyridoxal = ADP + pyridoxal 5'-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
4-amino-2-methyl-5-diphosphomethylpyrimidine biosynthesis (yeast)
-
-
Metabolic pathways
-
-
pyridoxal 5'-phosphate salvage I
-
-
pyridoxal 5'-phosphate salvage II (plants)
-
-
Vitamin B6 metabolism
-
-
vitamin B6 metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:pyridoxal 5'-phosphotransferase
Pyridoxine, pyridoxamine and various derivatives can also act as acceptors.
CAS REGISTRY NUMBER
COMMENTARY hide
9026-42-0
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
ecotype Columbia
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
male Mongolian gerbil
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
-
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
-
root growth of a sos4 mutant is significantly decreased when grown on either 100 mM or 200 mM sucrose as compared to root growth on10 mM sucrose. The sos4 mutant plant accumulates phytoglycogen
physiological function
additional information
-
experimental resurrection of the last common ancestor of the hydroxymethyl pyrimidine kinase group based on comparison of hydroxymethyl pyrimidine and pyridoxal kinases. Probably the last common ancestor was not able to use pyridoxal under physiological conditions. The pyridoxal kinase activity present in the current bifunctional enzymes must have appeared in a convergent event independently of the pyridoxal kinase activity of pdxY and pdxK genes. Substrate pyridoxal is 8-times less preferred than the phosphorylation of hydroxymethyl pyrimidine by the last ancestor
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
ATP + 4'-deoxypyridoxine
ADP + 4'-deoxypyridoxine 5'-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 4'-O-methylpyridoxine
ADP + 4'-O-methylpyridoxine 5'-phosphate
show the reaction diagram
-
-
-
-
?
ATP + 4-amino-5-hydroxymethyl-2-methylpyrimidine
ADP + 4-amino-5-phosphomethyl-2-methylpyrimidine
show the reaction diagram
ATP + 4-deoxypyridoxine
ADP + 4-deoxypyridoxine 5'-phosphate
show the reaction diagram
ATP + 5-deoxypyridoxal
ADP + ?
show the reaction diagram
ATP + ginkgotoxin
?
show the reaction diagram
-
ginkgotoxin is an effective pseudosubstrate
-
-
?
ATP + ginkgotoxin
ADP + ginkgotoxin 5'-phosphate
show the reaction diagram
-
-
-
-
?
ATP + omega-methylpyridoxal
ADP + omega-methylpyridoxal 5'-phosphate
show the reaction diagram
ATP + pyridoxal
ADP + pyridoxal 5'-phosphate
show the reaction diagram
ATP + pyridoxamine
ADP + pyridoxamine 5'-phosphate
show the reaction diagram
ATP + pyridoxine
ADP + pyridoxine 5'-phosphate
show the reaction diagram
methyl (1S)-1-[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylate + ATP
methyl (1S)-1-[3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylate + ADP
show the reaction diagram
-
-
very probably: methyl (1S)-1-[3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylate is an inhibitor of Plasmodium falciparum ornithine decarboxylase
-
?
methyl N-[[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]methyl]histidinate + ATP
methyl N-([3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]methyl)histidinate + ADP
show the reaction diagram
-
-
methyl N-([3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]methyl)histidinate is an inhibitor of Plasmodium falciparum ornithine decarboxylase: IC50 = 0.058 mM
-
?
methyl N-[[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]methyl]tryptophanate + ATP
methyl N-([3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]methyl)tryptophanate + ADP
show the reaction diagram
-
-
methyl N-([3-hydroxy-2-methyl-5-[(phosphonooxy)methyl]pyridin-4-yl]methyl)tryptophanate is an inhibitor of Plasmodium falciparum ornithine decarboxylase: IC50 = 0.064 mM
-
?
pyridoxal + ATP
pyridoxal 5'-phosphate
show the reaction diagram
-
-
-
-
?
pyridoxal + ATP
pyridoxal 5'-phosphate + ADP
show the reaction diagram
-
-
-
-
?
pyridoxamine + ATP
pyridoxamine 5'-phosphate
show the reaction diagram
-
-
-
-
?
pyridoxine + ATP
pyridoxine 5'-phosphate
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 + pyridoxal
ADP + pyridoxal 5'-phosphate
show the reaction diagram
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Li+
-
poor activator which seems to modify the enzymatic mechanism from a random to an ordered sequential pattern with ATP bound before pyridoxal. Km: 37 mM
Rb+
-
Km: 5.3 mM. Monovalent cation required, activation in the order of decreasing efficiency: K+, Rb+, NH4+
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(2-Diethylaminoethyl)hydrazine
-
0.005 mM, 20% inhibition
(R)-roscovitine
1,1-Bis(2-diethylaminoethyl)hydrazine
-
0.005 mM, 51% inhibition
1,4-diaminobutane
-
-
1,4-Diaminopiperazine
-
0.005 mM, 59% inhibition
1,6-diaminohexane
-
-
1,8-diaminooctane
-
-
1-(2-Hydrazinopropyl)piperidine
-
0.005 mM, 31% inhibition
1-(3-Dimethylamino-2-methylpropyl)hydrazine
-
0.005 mM, 39% inhibition
1-Aminopiperidine
-
0.005 mM, 58% inhibition
1-Cyclohexyl-1-methylhydrazine
-
0.005 mM, 29% inhibition
2,2'-hydrazinediethanol
-
0.005 mM, 47% inhibition
2,6-Dimethyl-3,4-bis(hydroxymethyl)pyridine
-
-
2-Methyl-3-amino-4,5,6-tris(hydroxymethyl)pyridine
-
-
2-Methyl-3-amino-4,5-bis(hydroxymethyl)pyridine
-
-
2-Methyl-3-hydroxy-5,6-bis(hydroxymethyl)pyridine
-
-
2-Methyl-4,5-bis(hydroxymethyl)pyridine
-
-
2-[1-(2-Diethylaminoethyl)hydrazine]ethanol
-
0.005 mM, 44% inhibition
3,4-dihydroxyphenylalanine
-
-
3-hydroxyanthranilic acid
-
IC50: 0.12 mM
3-hydroxykynurenine
-
IC50: 0.1 mM
4'-deoxypyridoxine
-
-
4'-O-methylpyridoxine
-
i.e. ginkgotoxin. Treatment leads to temporarily reduced pyridoxal phosphate formation in vitro and possibly in vivo
4-Aminomorpholine
-
0.005 mM, 52% inhibition
4-deoxypyridoxine
5-dimethylaminonaphthalene-1-sulfonyl-4-aminobutyrate
-
competitive with respect to pyridoxal
5-hydroxytryptamine
-
0.5 mM, 11% inhibition of activity with pyridoxine, 81% inhibition of activity with pyridoxal
alpha-Methylphenethylhydrazine
alpha-Methylphenethylhydrazone
aminooxyacetic acid
-
0.002 mM, 50% inhibition
azine
Ca2+
-
low substrate inhibition
cadaverine
-
-
Caffeine
-
21% inhibition at 0.1 mM
Chloroquine
about 40% inhibition at 1 mM
Cr2+
-
-
cycloserine
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 42% inhibition
D-penicillamine
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 20% inhibition
dopamine
enprofylline
-
33% inhibition at 0.1 mM
gamma-aminobutyric acid
-
-
ginkgotoxin
histamine
-
-
hydrazine
hydroxylamine
Isoniazid
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 81% inhibition
K+
-
at high concentrations
L-canaline
-
0.01 mM, 50% inhibition
lamotrigine
-
45% inhibition at 0.1 mM
levodopa
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 16% inhibition
Li+
-
at high concentrations
Mn2+
-
excess free divalent cations inhibit the enzyme
muzolimine
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 27% inhibition
N,N'-Bis(pyridoxyl)hydrazine
-
0.0005 mM, 50% inhibition
N-Dansyl-1,8-diaminooctane
NADH
-
0.5 mM, 14% inhibition of activity with pyridoxine, 11% inhibition of activity with pyridoxal
norepinephrine
-
-
Octylhydrazine
-
0.005 mM, 53% inhibition
Oxime
picolinate
-
0.5 mM, 95% inhibition of activity with pyridoxine, 97% inhibition of activity with pyridoxal
primaquine
progabide
-
0.1 mM, inhibits using either pyridoxamine or pyridoxal as substrate
putrescine
-
-
pyridoxal
pyridoxal 5'-phosphate
-
-
pyridoxal semicarbazone
-
0.00005 mM, 50% inhibition
pyridoxal-gamma-aminobutyrate
-
-
pyridoxaloxime
pyridoxamine
pyridoxine
-
substrate inhibition above 0.2 mM
quinolinate
-
0.5 mM, 82% inhibition of activity with pyridoxine, 82% inhibition of activity with pyridoxal
quinolinic acid
-
IC50: 0.42 mM
roscovitine
-
-
rugulactone
selectively modifies the enzyme not at the active site cysteine, but on a remote cysteine residue
-
Semicarbazine
-
0.0005 mM, 50% inhibition
serotonin
spermidine
-
weak
spermine
-
weak
Theobromine
-
22% inhibition at 0.1 mM
theophylline
Thiamphenicol
-
0.1 mM, inhibits activity with pyridoxal, but not with pyridoxamine as substrate, 31% inhibition
tryptamine
tyramine
-
-
xanthurenate
-
0.5 mM, 19% inhibition of activity with pyridoxine, 18% inhibition of activity with pyridoxal
xanthurenic acid
-
IC50: 0.36 mM
ZnATP2-
-
weak substrate inhibition
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
NAD+
-
0.5 mM, activation to 125%, activity with pyridoxine
phosphate
-
optimal concentration is 80-100 mM
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0216
4'-deoxypyridoxine
-
in 70 mM potassium phosphate buffer, pH 6.2; pH 6.2, 37C
0.00495
4'-O-methylpyridoxine
-
pH 6.2, 37C
1.85 - 2.03
4-Amino-5-hydroxymethyl-2-methylpyrimidine
0.0091 - 0.5
ATP
0.00495 - 0.0466
ginkgotoxin
0.17 - 0.42
MgATP
0.101 - 0.6
MgATP2-
0.194
MnATP2-
-
pH 5.8, 37C
0.00122 - 0.49
pyridoxal
0.006 - 0.126
pyridoxamine
0.00172 - 2.07
pyridoxine
0.06 - 0.07
ZnATP2-
additional information
additional information
-
effect of KCl on Km-values
-
TURNOVER NUMBER [1/s]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1.535 - 2.94
4'-deoxypyridoxine
0.46
4'-O-methylpyridoxine
Homo sapiens
-
pH 6.2, 37C
0.044 - 0.36
4-Amino-5-hydroxymethyl-2-methylpyrimidine
1.23 - 3.3
ATP
0.46 - 106
ginkgotoxin
0.015 - 131
pyridoxal
0.03 - 127
pyridoxamine
0.015 - 122
pyridoxine
kcat/KM VALUE [1/mMs-1]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
12.7 - 23.1
ATP
4
0.000228
ginkgotoxin
Trypanosoma brucei
-
recombinant enzyme, apparent value, in HEPES/MgCl2 pH 7.4, 0.15 mM ZnCl2, at 37C
6921
0.0009 - 127
pyridoxal
608
0.000214
pyridoxamine
Trypanosoma brucei
-
recombinant enzyme, apparent value, in HEPES/MgCl2 pH 7.4, 0.15 mM ZnCl2, at 37C
1284
0.000254
pyridoxine
Trypanosoma brucei
-
recombinant enzyme, apparent value, in HEPES/MgCl2 pH 7.4, 0.15 mM ZnCl2, at 37C
1051
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0574
4'-deoxypyridoxine
-
; pH 6.2, 37C
0.000414
4'-O-methylpyridoxine
-
pH 6.2, 37C
0.001
4-deoxypyridoxine
-
pH 6.0, 37C
0.002
5-dimethylaminonaphthalene-1-sulfonyl-4-aminobutyrate
-
pH 8.0, 37C
0.06
AMP
-
-
0.228
enprofylline
-
in 20 mM sodium BES buffer, pH 7.2, at 37C
0.000414 - 0.003
ginkgotoxin
0.056
lamotrigine
-
in 20 mM sodium BES buffer, pH 7.2, at 37C
1.54
Mg2+
-
recombinant enzyme, apparent value, in 70 mM potassium phosphate pH 7.0, 0.15 mM ZnCl2, at 37C
0.0572
primaquine
-
at pH 7.0 and 30C
0.0001
pyridoxal oxime
-
pH 7, 25C
0.002
pyridoxaloxime
-
pH 6.0, 37C
0.05
theophylline
-
in 20 mM sodium BES buffer, pH 7.2, at 37C
0.0398
Zn2+
-
recombinant enzyme, apparent value, in 70 mM potassium phosphate pH 7.0, 0.15 mM ZnCl2, at 37C
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.12
3-hydroxyanthranilic acid
Ovis aries
-
IC50: 0.12 mM
0.1
3-hydroxykynurenine
Ovis aries
-
IC50: 0.1 mM
0.42
quinolinic acid
Ovis aries
-
IC50: 0.42 mM
0.36
xanthurenic acid
Ovis aries
-
IC50: 0.36 mM
0.037
Zn2+
Homo sapiens
-
at pH 6.5 and 37C
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.004
-
brain enzyme
0.0125
0.02
-
substrate: methyl N-[[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]methyl]tryptophanate
0.038
-
substrate: methyl (1S)-1-[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]-2,3,4,9-tetrahydro-1H-beta-carboline-3-carboxylate
0.052
-
purified kinase 2 enzyme, pH 6.1, 37C
0.064
-
substrate: methyl N-[[3-hydroxy-5-(hydroxymethyl)-2-methylpyridin-4-yl]methyl]histidinate
0.07
-
crude enzyme, in 70 mM potassium phosphate (pH 5.5), 0.5 mM ZnCl2, at 37C
0.0713
-
-
0.1447
-
-
1.27
-
reaction with pyridoxal
1.42
-
reaction with pyridoxine
1.8
-
crude enzyme, in 70 mM potassium phosphate (pH 5.5), 0.5 mM ZnCl2, at 37C
2.105
-
-
2.47
-
-
2.5
-
purified enzyme, pH 6.1, 37C
2.7
-
purified kinase 1 enzyme, pH 6.1, 37C
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 6
5.8 - 6.3
-
-
8.8
-
enzyme from leukocytes
9.1
-
enzyme from erythrocytes
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4.5 - 8.5
-
the enzyme is inactive below pH 4.5. Enzyme activity decreases slowly above pH 6.0, to approximately 35% at pH 8.5
4.7 - 5.3
-
pH 4.7: about 60% of maximal activity, pH 5.3: about 40% of maximal activity
5 - 5.7
-
pH 5.0: about 90% of maximal activity, pH 5.7: about 45% of maximal activity
5 - 8
-
pH 5.0: about 70% of maximal activity, pH 8.0: about 50% of maximal activity
6 - 7
-
pH 6.0: about 50% of maximal activity, pH 7.0: about 50% of maximal activity
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
40
-
Zn2+ as activating anion
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.14
-
kinase 1, calculated from the deduced amino acid sequence
5.75
-
calculated from the deduced amino acid sequence
6.05
-
kinase 2, calculated from the deduced amino acid sequence
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
dentate granule cell. Pyridoxal kinase immunoreactivity is not increased after induction of long-term potentiation and not involved in increase in the efficiency of high frequency stimulus-induced potentiation of populations spike amplitude when compared with saline-, or Tat-protein-treated animals
Manually annotated by BRENDA team
-
from CA1 region, strong immunoreactivity
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
weak activity
Manually annotated by BRENDA team
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
splice variant SOS4.1 (but not SOS4.2) is localized in chloroplasts
Manually annotated by BRENDA team
PDB
SCOP
CATH
ORGANISM
UNIPROT
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Bacteroides thetaiotaomicron (strain ATCC 29148 / DSM 2079 / NCTC 10582 / E50 / VPI-5482)
Burkholderia multivorans (strain ATCC 17616 / 249)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Escherichia coli (strain K12)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Lactobacillus plantarum (strain ATCC BAA-793 / NCIMB 8826 / WCFS1)
Paraburkholderia xenovorans (strain LB400)
Pseudomonas aeruginosa (strain ATCC 15692 / DSM 22644 / CIP 104116 / JCM 14847 / LMG 12228 / 1C / PRS 101 / PAO1)
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
33000
-
x * 33000, SDS-PAGE
33100
-
x * 33100, calculated
33900
-
2 * 33900, SDS-PAGE
34861
-
x * 34861, calculation from nucleotide sequence
39500
-
2 * 39500, SDS-PAGE
57200
-
2 * 57200, calculated from the deduced amino acid sequence, 2 * 58000, SDS-PAGE, native mass by gel filtration
58000
-
2 * 57200, calculated from the deduced amino acid sequence, 2 * 58000, SDS-PAGE, native mass by gel filtration
60000
-
gel filtration, equilibrium sedimentation
65000
-
gel filtration
68000
-
gel filtration
73000
-
gel filtration
110000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hexamer
-
trimer or hexamer, X-ray crystallography
homodimer
monomer
trimer
-
trimer or hexamer, X-ray crystallography
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
hanging drop vapour diffusion method using 28% PEG 4000, 0.17 M sodium acetate trihydrate, and 0.1 M Tris-HCl (pH 8.5)
-
with 28% (w/v) PEG 4000, 0.17 M sodium acetate trihydrate, 0.1 M Tris-HCl (pH 8.5) as the precipitant in the presence of 10 mM ADP and 10 mM MgCl
-
hanging drop vapour diffusion method using 20 mM potassium phosphate, pH 7.5, 5 mM beta-mercaptoethanol, 0.2 mM EDTA, and 21% PEG 4K, 100 mM Tris-HCl, pH 8.5, 200 mM Na acetate, 40 mM MgSO4, and 10% glycerol
-
hanging drop vapour diffusion method using 0.1 mM Tris-HCl pH 8.0, 1.8 M NH4Ac, and 3% glycol
-
hanging drop vapour diffusion method using 100 mM Tris-HCl pH 8.0 and 50% 2-methyl-2,4-pentanediol; unliganded enzyme and in complex with MgATP, diffraction to 2.0 and 2.2 A rsolution, respectively. both Structures show similar open conformations. Mg2+ and Na+ act in tandem to anchor the ATP at the active site, which itself acts as a sink to bind several molecules of 2-methyl-2,4-pentanediol
-
in complex with ginkgotoxin and theophylline, hanging drop vapor diffusion method, using 48-50% (w/v) 2-methyl-1,3 propanediol as precipitant, at 22C
-
crystallized in the orthorhombic form using the hanging-drop vapour-diffusion method with sodium citrate as the precipitant, crystals are transferred into a soaking liquid without citrate and two-heavy-atom derivatives are prepared
-
crystals are grown in the presence of 100 mM potassium phosphate, pH 7.2, and 120 mM ammonium sulfate and crystals grown in the presence of 600 mM potassium phosphate, pH 7.2, and in the absence of ammonium sulfate. The crystals are quite stable to X-rays and diffract at 2.2 A resolution
-
hanging drop vapor diffusion method in complex with adenosine 5-(beta, gamma-methylenetriphosphate)-pyridoxamine, ADP-pyriodoxal 5-phosphate, and ADP
-
hanging drop vapor diffusion method, in complex with (R)-roscovitine and N6-methyl-(R)-roscovitine
sitting drop vapor diffusion method, using 0.1 M Tris, pH 8.5, 1 M LiCl2, and 15% (w/v) PEG 6000
native protein and in complex with its substrates to 1.4?1.85 A resolution. The protein shows a typical ribokinase fold with a central large beta-sheet consisting of nine strands, flanked by three and five structurally conserved alpha-helices
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 11
-
25C, 3 h, stable. Gradual irreversible inactivation below pH 5.5
641425
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
-20
-
stable at -20C for approximately 21 days
25
-
pH 5.5-11, 3 h, stable
40
-
greatest stability is at below 40C
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
3 M guanidine hydrochloride, enzyme loses more than 90% of the alpha-helix content
-
if instead of the phosphate buffer an acetate buffer is used, enzymatic activity is reduced to 74% and almost no activity is recorded when a citrate buffer is used
-
impure enzyme is stabilized by Zn2+
-
stable for 48 h when transduced into PC-12 cells
-
the enzyme is digested by chymotrypsin to proteolytic fragments of 24000 Da and 16000 Da
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-18C, stable for 20 weeks
-
-20C, recombinant enzyme, stable for 21 days
-
-80C, 20 mM phosphate buffer, pH 7.3, stable for at least some weeks
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
from hippocampal tissue
-
Hi-Load Superdex 200 gel filtration
-
immobilized metal ion adsorption chromatography (Ni2+)
-
Ni Sepharose affinity column chromatography
-
Ni-NTA agarose column chromatography
Ni-NTA agarose column chromatography and HiLoad 26/60 Superdex 200 gel filtration
-
Ni-NTA column chromatography
-
Ni-NTA column chromatography and Mono Q column chromatography
Ni-NTA Sepharose column chromatography
-
Phenyl ToyoPearl 650S column chromatography and Hi-Load Superdex 200 column gel filtration
-
pyridoxyl-EAH-Sepharose 4B column chromatography; recombinant enzyme
-
Q-Sepharose column chromatography and Superdex 75 gel filtration
-
recombinant enzyme
recombinant protein using His-tag
-
recombinant protein using Strep-tag
-
TMAE column chromatography, Phenyl Sepharose column chromatography, and hydroxyapatite column chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
; expressed in Escherichia coli strain BL21 (DE3)
-
expressed as His-tag fusion protein in Escherichia coli BL21(DE3)
-
expressed as Strep-tag fusion protein in Escherichia coli BLR(DE3)
-
expressed in Escherichia coli
expressed in Escherichia coli BL21 (DE3) cells
-
expressed in Escherichia coli BL21(DE3) cells
expressed in Escherichia coli HMS174(DE3)
-
expressed in Escherichia coli Rosetta (DE3) cells
-
expressed in Escherichia coli Rosetta (DE3)pLysS cells; expression in Escherichia coli
-
expressed in Escherichia coli strain BL21 (DE3) plysS
-
expression in Escherichia coli
His-tag, expressed in Escherichia coli Rosetta (lambdaDE3)
-
kinase 1 and 2 expressed in Escherichia coli HMS174(DE3), kinase 2 additionally expressed as His-tag fusion protein
-
recombinant enzyme is overexpressed in Escherichia coli as a fusion protein with maltose binding protein
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
expression patterns show that the gene is involved in abiotic stress and phytohormone regulation; expression patterns show that the gene is involved in abiotic stress and phytohormone regulation
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
D235A
-
active site residue
D235N
-
active site residue
C124A
the mutant shows about 2fold increased catalytic efficiency compared to the wild type enzyme
C124A
-
the mutant shows about 2fold increased catalytic efficiency compared to the wild type enzyme
-
D225A
-
inactive
-
C110A
residue C110 is mandatory for pyridoxal, but not for pyridoxine, or 4-amino-5-hydroxymethyl-2-methylpyrimidine turnover
C214A
mutant does not turn over pyridoxal, pyridoxine, or 4-amino-5-hydroxymethyl-2-methylpyrimidine. Residue C214 acts as the catalytic base in the phosphorylation reaction
C110A
-
residue C110 is mandatory for pyridoxal, but not for pyridoxine, or 4-amino-5-hydroxymethyl-2-methylpyrimidine turnover
-
C214A
-
mutant does not turn over pyridoxal, pyridoxine, or 4-amino-5-hydroxymethyl-2-methylpyrimidine. Residue C214 acts as the catalytic base in the phosphorylation reaction
-
additional information
plants lacking enzyme activity display chlorosis and reduced plant size, and hypersensitivity to NaCl and sucrose. Mutant has higher level of pyridoxine, pyridoxamine, and pyridoxal 5'-phosphate. Mutant shows higher activity of pyridoxine/pyridoxamine 5'-phosphate oxidase as well as of the vitamin B6 de novo pathway enzyme Pdx1 and increased total vitamin B6 levels
Renatured/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
the enzyme which has lost more than 90% of the alpha-helix content after treatment with 3 M guanidine hydrochloride regains more than 90% of the original catalytic activity after overnight dialysis against 10 mM potassium phosphate, pH 7 at 4C
-
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
agriculture
plants lacking enzyme activity display chlorosis and reduced plant size, and hypersensitivity to NaCl and sucrose. Mutant has higher level of pyridoxine, pyridoxamine, and pyridoxal 5'-phosphate. Mutant shows higher activity of pyridoxine/pyridoxamine 5'-phosphate oxidase as well as of the vitamin B6 de novo pathway enzyme Pdx1 and increased total vitamin B6 levels
medicine
Show AA Sequence (6111 entries)
Longer loading times are possible. Please use the Sequence Search for a specific query.