Information on EC 2.7.1.138 - ceramide kinase

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The enzyme appears in viruses and cellular organisms

EC NUMBER
COMMENTARY hide
2.7.1.138
-
RECOMMENDED NAME
GeneOntology No.
ceramide kinase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
ATP + ceramide = ADP + ceramide 1-phosphate
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
phospho group transfer
-
-
-
-
Phosphorylation
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
sphingolipid biosynthesis (plants)
-
-
Sphingolipid metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
ATP:ceramide 1-phosphotransferase
-
CAS REGISTRY NUMBER
COMMENTARY hide
123175-68-8
-
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
metabolism
-
sphingolipid metabolism
physiological function
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
alpha-OH-C24:0 phytoceramide + ATP
alpha-OH-C24:0 phytoceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
alpha-OH-D-erythro-C6:0 ceramide + ATP
alpha-OH-D-erythro-C6:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
ATP + 3-O-methyl C16:0 ceramide
ADP + 3-O-methyl C16:0 ceramide 1-phosphate
show the reaction diagram
-
very low activity
-
-
?
ATP + C16-ceramide
ADP + C16-ceramide 1-phosphate
show the reaction diagram
ATP + C18-ceramide
ADP + C18-ceramide 1-phosphate
show the reaction diagram
-
lower activity
-
-
?
ATP + C2-ceramide
ADP + C2-ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + C8-ceramide
ADP + C8-ceramide 1-phosphate
show the reaction diagram
-
best substrate
-
-
?
ATP + C8-dihydroceramide
ADP + C8-dihydroceramide 1-phosphate
show the reaction diagram
-
high activity
-
-
?
ATP + C8-phytoceramide
ADP + C8-phytoceramide 1-phosphate
show the reaction diagram
-
high activity
-
-
?
ATP + ceramide
ADP + ceramide 1-phosphate
show the reaction diagram
ATP + D-erythro-C6:0 ceramide
ADP + D-erythro-C6:0 ceramide 1-phosphate
show the reaction diagram
-
very low activity
-
-
?
ATP + D-erythro-N-hexanoyl-sphinganine
ADP + D-erythro-N-hexanoyl-sphinganine 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-erythro-N-hexanoyl-sphingenine
ADP + D-erythro-N-hexanoyl-sphingenine 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C10:0 ceramide
ADP + D-eythro-C10:0 ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C12:0 ceramide
ADP + D-eythro-C12:0 ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C14:0 ceramide
ADP + D-eythro-C14:0 ceramide 1-phosphate
show the reaction diagram
-
best substrate
-
-
?
ATP + D-eythro-C16:0 (R)-alpha-hydroxyceramide
ADP + D-eythro-C16:0 (R)-alpha-hydroxyceramide 1-phosphate
show the reaction diagram
-
low activity
-
-
?
ATP + D-eythro-C16:0 (S)-alpha-hydroxyceramide
ADP + D-eythro-C16:0 (S)-alpha-hydroxyceramide 1-phosphate
show the reaction diagram
-
very low activity
-
-
?
ATP + D-eythro-C16:0 ceramide
ADP + D-eythro-C16:0 ceramide 1-phosphate
show the reaction diagram
ATP + D-eythro-C16:0 dehydroceramide
ADP + D-eythro-C16:0 dehydroceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C16:0 dihydroceramide
ADP + D-eythro-C16:0 dihydroceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C16:0 urea ceramide
ADP + D-eythro-C16:0 urea ceramide 1-phosphate
show the reaction diagram
-
low activity, no activity with L-e-C16:0 urea ceramide
-
-
?
ATP + D-eythro-C16:0-cis ceramide
ADP + D-eythro-C16:0-cis ceramide 1-phosphate
show the reaction diagram
-
low activity
-
-
?
ATP + D-eythro-C18:0 ceramide
ADP + D-eythro-C18:0 ceramide 1-phosphate
show the reaction diagram
ATP + D-eythro-C18:0 phytoceramide
ADP + D-eythro-C18:0 phytoceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C20:0 ceramide
ADP + D-eythro-C20:0 ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C24:0 ceramide
ADP + D-eythro-C24:0 ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C24:1 ceramide
ADP + D-eythro-C24:1 ceramide 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + D-eythro-C8:0 ceramide
ADP + D-eythro-C8:0 ceramide 1-phosphate
show the reaction diagram
-
low activity
-
-
?
ATP + diacylglycerol
ADP + diacylglycerol 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + hexadecanoylceramide
ADP + hexadecanoylceramide 1-phosphate
show the reaction diagram
C8 acyl chain, slightly higher activity than with natural ceramide
-
?
ATP + hexanoylceramide
ADP + hexanoylceramide 1-phosphate
show the reaction diagram
C6 acyl chain, similar activity than with natural ceramide, very low activity with C2-ceramide and C6-dihydroceramide, no activity with C2-dihydroceramide
-
?
ATP + N-acetyl-D-erythro-sphingenine
ADP + N-acetyl-D-erythro-sphingenine 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-acetyl-sphingenine
ADP + N-acetyl-sphingenine 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-hexadecanoyl-sphingenine
ADP + N-hexadecanoyl-sphingenine 1-phosphate
show the reaction diagram
ATP + N-hexanoyl-1-O-hexadecyl-2-deoxy-2-amino-sn-glycerol
ADP + ?
show the reaction diagram
-
-
-
-
?
ATP + N-hexanoyl-D-erythro-sphingenine
ADP + N-hexanoyl-D-erythro-sphingenine 1-phosphate
show the reaction diagram
-
best substrate
-
-
?
ATP + N-hexanoyl-sphingenine
ADP + N-hexanoyl-sphingenine 1-phosphate
show the reaction diagram
-
-
-
-
?
ATP + N-tetradecanoyl-(2S)-amino-butan-1-ol
ADP + ?
show the reaction diagram
-
-
-
-
?
ATP + N-tetradecanoyl-(2S)-amino-hexadecan-1-ol
ADP + ?
show the reaction diagram
-
-
-
-
?
ATP + N-tetradecanoyl-2-amino-1-butanol
ADP + ?
show the reaction diagram
-
-
-
-
?
ATP + N-tetradecanoyl-2S-amino-1-butanol
ADP + ?
show the reaction diagram
-
-
-
-
?
ATP + octanoylceramide
ADP + octanoylceramide 1-phosphate
show the reaction diagram
C8 acyl chain, higher activity than with natural ceramide
-
?
ATP + octanoyldihydroceramide
ADP + octanoyldihydroceramide 1-phosphate
show the reaction diagram
C8 acyl chain, slightly higher activity than with natural ceramide
-
?
ATP + stearoylceramide
ADP + stearoylceramide 1-phosphate
show the reaction diagram
-
-
-
?
C12-ceramide + ATP
C12-ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
C16-ceramide + ATP
C16-ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
C2-ceramide + ATP
C2-ceramide 1-phosphate + ADP
show the reaction diagram
C26:0 phytoceramide + ATP
C26:0 phytoceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
C8-ceramide + ATP
C8-ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
ceramide + ATP
ceramide 1-phosphate + ADP
show the reaction diagram
D-erythro-C12:0 ceramide + ATP
D-erythro-C12:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-C14:0 ceramide + ATP
D-erythro-C14:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-C16:0 ceramide + ATP
D-erythro-C16:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
used for kinetic analysis of ceramide kinase activity
-
-
?
D-erythro-C2:0 ceramide + ATP
D-erythro-C2:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-C2:0 dihydroceramide + ATP
D-erythro-C2:0 dihydroceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-C6:0 ceramide + ATP
D-erythro-C6:0 ceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-C6:0 dihydroceramide + ATP
D-erythro-C6:0 dihydroceramide 1-phosphate + ADP
show the reaction diagram
-
-
-
-
?
D-erythro-N-hexanoyl-sphingenine + ATP
D-erythro-N-hexanoyl-sphingenine 1-phosphate + ADP
show the reaction diagram
-
D-erythro isomer of C6-ceramide, the specific CERK activity in cells exposed for 16-18 h to 0.005 mM C6-ceramide dropped to 18.7%
-
-
?
D-threo-C6-ceramide + ATP
D-threo-C6-ceramide 1-phosphate + ADP
show the reaction diagram
-
no or only weakly decrease of the specific CERK activity
-
-
?
GTP + ceramide
GDP + ceramide 1-phosphate
show the reaction diagram
-
26% of the activity with ATP at 1 mM
-
?
L-erythro-C6-ceramide + ATP
L-erythro-C6-ceramide 1-phosphate + ADP
show the reaction diagram
-
no or only weakly decrease of the specific CERK activity
-
-
?
L-threo-C6-ceramide + ATP
L-threo-C6-ceramide 1-phosphate + ADP
show the reaction diagram
-
no or only weakly decrease of the specific CERK activity
-
-
?
N-((4-(4,4-difluoro-5-(2-thienyl)-4-bora-3alpha,4alpha-diaza-s-indacene-3-yl)phenoxy)acetyl)sphingosine + ATP
N-((4-(4,4-difluoro-5- (2-thienyl)-4- bora-3a,4a-diaza-s-indacene-3-yl)phenoxy)acetyl)sphingosine 1-phosphate + ADP
show the reaction diagram
-
TRB-ceramide
-
-
?
N-(12-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)dodecanoyl)-D-erythro-sphingosine + ATP
N-(12-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)dodecanoyl)-D-erythro-sphingosine 1-phosphate + ADP
show the reaction diagram
-
C12-NBD ceramide
-
-
?
N-(5-(5,7-dimethyl-BODIPY)-L-pentanoyl)-D-erythrosphingosine + ATP
N-(5-(5,7-dimethyl-BODIPY)-L-pentanoyl)-D-erythrosphingosine 1-phosphate + ADP
show the reaction diagram
-
DMB-C5 ceramide
-
-
?
N-(6-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)hexanoyl)-D-erythro-sphingosine + ATP
N-(6-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)hexanoyl)-D-erythro-sphingosine 1-phosphate + ADP
show the reaction diagram
N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl-D-erythro-sphingosine + ATP
N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl-D-erythro-sphingosine 1-phosphate + ADP
show the reaction diagram
-
NBD-ceramide
NBD-ceramide 1-phosphate
-
?
N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl-D-erythrosphingosine + ATP
N-(7-(4-nitrobenzo-2-oxa-1,3-diazole))-6-aminocaproyl-D-erythrosphingosine 1-phosphate + ADP
show the reaction diagram
N-(8-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)octanoyl)-D-erythro-sphingosine + ATP
N-(8-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)octanoyl)-D-erythro-sphingosine 1-phosphate + ADP
show the reaction diagram
-
C8-ceramide
-
-
?
N-hexanoyl-sphinganine + ATP
N-hexanoyl-sphinganine 1-phosphate + ADP
show the reaction diagram
-
C6-dihydroceramide
-
-
?
N-octyl-sphingenine + ATP
N-octyl-sphingenine 1-phosphate + ADP
show the reaction diagram
-
C8-ceramine, no or only weakly decrease of the specific CERK activity
-
-
?
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 + C16-ceramide
ADP + C16-ceramide 1-phosphate
show the reaction diagram
ATP + ceramide
ADP + ceramide 1-phosphate
show the reaction diagram
ATP + N-hexadecanoyl-sphingenine
ADP + N-hexadecanoyl-sphingenine 1-phosphate
show the reaction diagram
-
-
-
-
?
ceramide + ATP
ceramide 1-phosphate + ADP
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
no activity with GTP
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
NaCl
-
at 50 mM
additional information
-
no stimulation by Mn2+, Ba2+, Cd2+ and Zn2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
(4aR,6aS,12aS,12bR)-10-hydroxy-4,4,6a,12b-tetramethyl-1,3,4,4a,5,6,6a,12,12a,12b-decahydro-2H-benzo(a)xanthene-8,11-dione
-
KD, enantiomer of K1, strongly inhibits CerK activity
(4aS,6aR,12aR,12bS)-10-hydroxy-4,4,6a,12b-tetramethyl-1,3,4,4a,5,6,6a,12,12a,12b-decahydro-2H-benzo(a)xanthene-8,11-dione
1alpha,25-dihydroxyvitamin D3
-
enzyme activity decreases by approximately 40% in the presence of the hormone
2,5-dihydroxy-3-(((1R,4aR,8aR)-5,5,8a-trimethyl-2-methylidenedecahydronaphthalen-1-yl)methyl)cyclohexa-2,5-diene-1,4-dione
-
KB, enantiomer of K1, strongly inhibits CerK activity
2,5-dihydroxy-3-(((1S,2S,4aS,8aR)-2,5,5,8a-tetramethyldecahydronaphthalen-1-yl)methyl)cyclohexa-2,5-diene-1,4-dione
-
KC, F12509A with an additional hydrogen at the double bond in methylene position, weak inhibition
2,5-dihydroxy-3-(((1S,4aS,8aS)-5,5,8a-trimethyl-2-methylidenedecahydronaphthalen-1-yl)methyl)cyclohexa-2,5-diene-1,4-dione
-
F12509A, inhibitor of sphingosine kinase 1, weak inhibition
2,5-dihydroxy-3-[[(4aS,8aS)-2,5,5,8a-tetramethyl-3,4,4a,5,6,7,8,8a-octahydronaphthalen-1-yl]methyl]cyclohexa-2,5-diene-1,4-dione
-
K1, novel F-12509A olefin isomer, inhibitor decreases ceramide 1-phosphate levels without changing other lipids such as ceramide and sphingomyelin
3,3-cholamidopropyl-dimethylammonio-1-propanesulfonate
-
almost complete inhibition at high concentrations
3-(cyclohexylmethyl)-2,5-dihydroxycyclohexa-2,5-diene-1,4-dione
-
K2, cyclohexane derivative with a benzoquinone, weak inhibition
adamantane-1-carboxylic acid (2-benzoylamino-benzothiazol-6-yl)amide
-
NVP-231, a potent, specific, and reversible CerK inhibitor that competitively inhibits binding of ceramide to CerK
all-trans retinoic acid
-
inhibits ceramide kinase transkription in neuroblastoma cells
C2-ceramide
-
-
Ca2+
-
above 1 mM; inhibition above 1 mM
D-erythro-N,N-dimethylsphingosine
-
-
dimethylsphingosine
-
DMS
F-12509A
-
0.1 mM, 40-50% inhibition, sphingosine kinase inhibitor
KN-93
-
-
lauryldimethylammonium N-oxide
-
; inhibitory at low concentrations
N,N-dimethylsphingosine
-
0.1 mM, 65% inhibition
N-acetyl-D-erythro-sphingenine
-
substrate inhibition above 0.1 mM
N-acetylsphingosine
-
i.e. C2-Cer, in vivo during inhibition of mast cell degranulation
N-ethyl-maleimide
-
thiol modifiying agent stops CerK acitivity, demonstrating that CerK contains exposed cysteine residues important for enzymatic activity
N-laurylsarcosine
-
inhibitory at low concentrations
NVP-231
octyl-beta-D-glucopyranoside
-
almost complete inhibition at high concentrations
siRNA
-
sphinganine
-
-
sphingenine
-
-
sphingosine
-
0.1 mM, 55% inhibition
W-7
-
calmodulin antagonist
ZK191784
-
enzyme activity decreases by approximately 25% in the presence of the hormone analogue
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
A23187
-
calcium ionophore, 216% activation of ceramide kinase in A549 lung adenocarcinoma cells
Ca2+
-
1 mM, stimulatory effect in the absence of Mg2+
Calcium
-
-
Calmodulin
-
binding site is located at residues 422-435, which form a amphipathic helical wheel, motif 1-8-14, calmodulin is involved in the Ca2+-dependent activation of ceramide kinase as a calcium-sensor
interleukin-1 beta
-
187% activation of ceramide kinase in A549 lung adenocarcinoma cells
-
Mg2+
-
3 mM, highly dependent on Mg2+
peroxisome proliferator-activated receptor beta
-
upregulation of CERK through activation of peroxisome proliferator-activated receptor beta (PPARbeta) results in decreased intracellular ceramide levels and increased cell survival with less cell death
-
Phorbol esters
-
e.g. PMA
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.025 - 85.6
ATP
0.107
C12-ceramide
-
-
0.045 - 20.9
ceramide
0.001
N-(6-((7-nitro-2-1,3-benzoxadiazol-4-yl)amino)hexanoyl)-D-erythro-sphingosine
-
-
0.02
N-acetyl-D-erythro-sphingenine
-
pH 7.2, 37C, recombinant enzyme
0.022
N-acetyl-sphingenine
-
-
0.03
N-hexanoyl-D-erythro-sphingenine
-
pH 7.2, 37C, recombinant enzyme
0.03
N-hexanoyl-sphingenine
-
-
0.004 - 0.009
stearoylceramide
additional information
additional information
-
kinetics of recombinant enzymes
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0033
(4aR,6aS,12aS,12bR)-10-hydroxy-4,4,6a,12b-tetramethyl-1,3,4,4a,5,6,6a,12,12a,12b-decahydro-2H-benzo(a)xanthene-8,11-dione
-
-
0.0022
(4aS,6aR,12aR,12bS)-10-hydroxy-4,4,6a,12b-tetramethyl-1,3,4,4a,5,6,6a,12,12a,12b-decahydro-2H-benzo(a)xanthene-8,11-dione
-
-
0.452
2,5-dihydroxy-3-(((1S,2S,4aS,8aR)-2,5,5,8a-tetramethyldecahydronaphthalen-1-yl)methyl)cyclohexa-2,5-diene-1,4-dione
-
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.000012
adamantane-1-carboxylic acid (2-benzoylamino-benzothiazol-6-yl)amide
Homo sapiens
-
-
0.000012
NVP-231
Homo sapiens
-
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.000000004
-
liver of wild-type mice
0.000000028
-
kidney of wild-type mice
0.000000038
-
pancreas of wild-type mice
0.000001191
-
testis of wild-type mice
0.00000141
-
cerebellum of wild-type mice
0.00125
-
pH 7.2, 30C, recombinant ceramide kinase
0.92 - 5.4
-
recombinant enzyme in CHO cell extract, substrate N-C6-D-erythro-sphingenine
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6.2 - 7.8
-
-
6.5 - 7
-
-
pH RANGE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
5.5 - 8
-
approx. 20% of maximal activity at pH 5.5 and pH 8.0
6 - 8
-
approx. 25% of maximal activity at pH 6.0 and pH 8.0
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
extraction of bone marrow and preparation of macrophages
Manually annotated by BRENDA team
-
mouse keratinocyte cell line SP1
Manually annotated by BRENDA team
-
cell line HL-60
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
leukocyte
Manually annotated by BRENDA team
-
very high expression level in resting and low expression level in activated CD4+ cell, resting CD8+ cells, resting, CD14+ cells, higher expression level in resting and low expression level in activated CD19+ cells
Manually annotated by BRENDA team
-
bone marrow-derived macrophage, BMDM
Manually annotated by BRENDA team
-
CD4+ and CD8+
Manually annotated by BRENDA team
-
low expression level
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
CERK and localizes to the caveolar microdomains during phagocytosis
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
60000
-
SDS-PAGE
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
lipoprotein
-
the enzyme is myristoylated at the N-terminus within the pleckstrin homology PH domain, required for activity
phosphoprotein
-
phosphorylation of serine residues at positions 340 and 408, putative additional phosphorylation site at serine 427 lying in the calcium/calmodulin domain, preventing phosphorylation at S340 results in CERK instability
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
20% glycerol and 0.05 mM ATP stabilize during solubilization
-
solubilized enzyme loses activity overnight at 4C
-
OXIDATION STABILITY
ORGANISM
UNIPROT
LITERATURE
ceramide kinase is extremely sensitive to oxidation, 1 mM dithiothreitol stabilizes
-
640967
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-70C, 48 h, 50% loss of activity
-
30C, 50 mM Tris/HCl, pH 7.5, 270 mM sucrose, 1 mM EDTA, 1 mM EGTA, 5 mM tris(hydroxypropyl)phosphine, 1% Triton X-100, 20 min, 50% decrease of activity
-
4C, solubilized, 1 week
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Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
anti-FLAG M2 affinity gel chromatography
-
as a C-terminal fusion to maltose binding protein, MBP-Cerk was purified on amylose resin
-
by affinity chromatography
-
Ni-NTA affinity column chromatography
-
nickel-nitriloacetate resin chromatography
-
partially, subcellular fractionation
Sepharose-bead chromatography and Bio-Spin 6 column chromatography
-
Triton X-114, Mono Q
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Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
DNA and amino acid sequence determination, expression in HeLa and COS-1 cells, expression of GFP-tagged enzyme in HUVEC and COS-1 cells
-
expressed in CHO cells
-
expressed in CHO cells; expressed in Escherichia coli Top10F cells; expression in Escherichia coli strain BL21(DE3), expression in CHO cells
-
expressed in COS-1 cells
-
expressed in COS-1 cells; expressed in COS-1 fibroblasts and RBL-2H3 basophils; expression in Escherichia coli, overexpression of human CerK in COS-1 and RBL-2H3 cells
-
expressed in COS-1 cells; recombinant full-length hCERK is obtained from baculovirus-infected Sf9 cells as a glutathione-S-transferase fusion
-
expressed in COS-7 cells
-
expressed in J-774 cells; expression in Escherichia coli
-
expression in baculovirus-infected SF9 insect cells
-
expression in baculovirus-infected Sf9, overexpression in COS1-CerK cells
-
expression in CHO cells in the plasma membrane
-
expression in SF9 insect cell and transfection of FLAG-tagged or GFP-tagged wild type and mutant CerK proteins in COS-1 cell
-
expression of C-terminally FLAG-tagged and of EGFP-tagged wild-type and mutant enzymes in COS-1 cells
-
expression of green fluorescent protein-tagged human ceramide kinase in CHO-W11A cells, the increase in ceramide 1-phosphate formation by transfection with the vector for human ceramide kinase significantly enhances the Ca2+-ionophore A23187-induced release of arachidonic acid via cytosolic phospholipase A2alpha activation in CHO cells
-
expression of hCERK in COS-1 cells
-
expression of wild-type and mutant enzymes in rat basophilic leukemia RBL-2H3 cells, expression of wild-type and mutant enzymes in CHO cells
-
overexpression in HeLa cells, expression of His6-tagged enzyme in Spodoptera frugiperda Sf9 insect cells using the baculovirus infection system
-
stable expression in the cytosol and plasma membrane of rat basophilic leukemia RBL-2H3 cells, overexpression of the enzyme enhances the level of degranulation
-
stable expression of human CerK in COS-1 cell, named COS-CerK cell, and RBL-2H3 cell
-
transient expression in HEK293 cells
-
transient overexpression of FLAG-tagged wild-type and mutant enzymes in HEK293 cells
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
all-trans retinoic acid inhibits ceramide kinase transkription during neuronal differentiation of a human neuroblastoma cell line SH-SY5Y, inhibition of the transcriptional activity of the 5'-promoter of ceramide kinase. Retinol does not reduce CERK protein and mRNA level
-
ceramide kinase expression shows no association with age, lymph node status and tumor size
-
Cerk is rapidly upregulated in tumor cells following HER2/neu downregulation or treatment with adriamycin
-
DCERK protein is expressed during all developmental stages
-
expression of the enzyme, after 24 h of treatment with 1alpha,25-dihydroxyvitamin D3 or its analogue ZK191784, is reduced by approximately 40 and 60%, respectively
-
higher expression in estrogen receptor negative breast tumors, gene expression of ceramide kinase is associated with prognosis in breast cancer, high ceramide kinase expression is correlated with ErbB2 status
-
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C324A
-
same specific acitity as wild-type CerK
C398A
-
same specific acitity as wild-type CerK
CERKL
-
the point mutant enzyme CERKL does not phosphorylate ceramide or diacylglycerol and is localized in the nucleus
DELTA1-123
-
production by site-directed mutagenesis, localization in cytosol
DELTA124-537
-
production by site-directed mutagenesis, resulted in almost complete accumulation in the nucleus
DELTA219-496
-
production by site-directed mutagenesis, localization in cytosol and nucleus
DELTA340-537
-
production by site-directed mutagenesis, significant accumulation into the nucleus, existence of nuclear export signals in the C-terminal part of enzyme, traditional nuclear export signals 511-IEVRVHCQLVRL-522 in the CC3 domain and a class 2 nuclear export signals 347-CRAGCFVC-354 between the CC1 and the CC2 domains
DELTA454-537
-
production by site-directed mutagenesis, localization in cytosol and nucleus
DELTA514-537
-
production by site-directed mutagenesis, nuclear localization of ceramide kinase
DELTA520-537
-
production by site-directed mutagenesis, localization is mostly cytosolic but is also detected in the nucleus
DELTA525-537
-
production by site-directed mutagenesis, both localization and cellular activity are lost
DELTA528-537
-
production by site-directed mutagenesis, no effect of localization and activity when assaying at the cellular level with exogenously added substrate, activity is lost after cell lysis in vitro
DELTA533-537
-
production by site-directed mutagenesis, no effect of localization and activity
G2A
-
site-directed mutagenesis, reduced activity compared to the wild-type enzyme
K68A/K74A/K80A
-
shows wild type activity
K90V/K98V
-
shows partial activity
K90V/R91A
-
shows partial activity
K90V/R91A/R96A/K98V
-
totally devoid of activity
R29A/R33A/R36A
-
shows wild type activity
S340A
-
by site directed mutagenesis, using of Triton X-100 as lysis buffer results in 15% activity decrease in the mutant protein compared with the wild type enzyme, when octylglucoside is used instead of Triton X-100 for cell lysis, activity is reduced in the S340A mutant protein which reaches only 15% of wild type activity
S340D
-
by site directed mutagenesis, intermediate recovery of 16% is observed for the mutant protein
S427A
-
by site directed mutagenesis, activity in the S427A mutant protein amounts to only 30% of that of wild type enzyme
S427D
-
by site directed mutagenesis, mutant protein displays 50% of wild type activity
E8A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
F429R
-
site-directed mutagenesis, weak binding of calmodulin compared to the wild-type enzyme
F431R
-
site-directed mutagenesis, no binding of calmodulin in contrary to the wild-type enzyme
G2A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
L10A
-
site-directed mutagenesis, 99% reduced activity compared to the wild-type enzyme, substrate affinity and activation by Ca2+ are unaffected, mutation within the pleckstrin homology domain
L10I
-
site-directed mutagenesis, 71% reduced activity compared to the wild-type enzyme, substrate affinity and activation by Ca2+ are unaffected, mutation within the pleckstrin homology domain
L422R
-
site-directed mutagenesis, similar binding of calmodulin compared to the wild-type enzyme
L422R/F429R
-
site-directed mutagenesis
L435R
-
site-directed mutagenesis, no binding of calmodulin in contrary to the wild-type enzyme
P9A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
S12A
-
site-directed mutagenesis, the mutant shows unaltered activity compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
analysis
-
enzymatic assay to quantify mass levels of long chain ceramides in cellular lipid extracts
diagnostics
-
putative prognostic marker in breast cancer, estrogen receptor negative patients with high ceramide kinase expression had a worse prognosis then those with low expression
medicine