Information on EC 2.3.1.43 - phosphatidylcholine-sterol O-acyltransferase

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

EC NUMBER
COMMENTARY hide
2.3.1.43
-
RECOMMENDED NAME
GeneOntology No.
phosphatidylcholine-sterol O-acyltransferase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
phosphatidylcholine + a sterol = 1-acylglycerophosphocholine + a sterol ester
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Acyl group transfer
-
-
-
-
transesterification
-
-
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Glycerophospholipid metabolism
-
-
SYSTEMATIC NAME
IUBMB Comments
phosphatidylcholine:sterol O-acyltransferase
Palmitoyl, oleoyl and linoleoyl residues can be transferred; a number of sterols, including cholesterol, can act as acceptors. The bacterial enzyme also catalyses the reactions of EC 3.1.1.4 phospholipase A2 and EC 3.1.1.5 lysophospholipase.
CAS REGISTRY NUMBER
COMMENTARY hide
9031-14-5
-
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
goat
-
-
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
horse
-
-
Manually annotated by BRENDA team
chicken
Uniprot
Manually annotated by BRENDA team
-
-
-
Manually annotated by BRENDA team
C57BL/6 mice
-
-
Manually annotated by BRENDA team
pig
-
-
Manually annotated by BRENDA team
gene lcat
UniProt
Manually annotated by BRENDA team
gene lcat
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
evolution
malfunction
metabolism
physiological function
additional information
-
high plasma lecithin:cholesterol acyltransferase activity does not predict low incidence of cardiovascular events, a possible attenuation of cardioprotection is associated with high HDL cholesterol
SUBSTRATE
PRODUCT                       
REACTION DIAGRAM
ORGANISM
UNIPROT
COMMENTARY
(Substrate) hide
LITERATURE
(Substrate)
COMMENTARY
(Product) hide
LITERATURE
(Product)
Reversibility
r=reversible
ir=irreversible
?=not specified
1,2-bis-(1-pyrenebutanoyl)-sn-glycero-3-phosphocholine + cholesterol
cholesteryl-1-pyrenebutyrate + lysophosphatidylcholine
show the reaction diagram
1,2-bis[4-(1-pyreno)butanoyl]-sn-glycero-3-phosphocholine + cholesterol
?
show the reaction diagram
fluorescent substrate assay
-
-
?
1-acylglyceryl phosphorylcholine + lecithin
lecithin + 1-acylglyceryl phosphorylcholine
show the reaction diagram
-
transfer of an acyl group from a lecithin molecule to another on the low-density lipoprotein surface, lysolecithin acyltransferase activity
-
r
1-O-alkyl-2-acetyl-sn-glycerol-3-phosphocholine + 1-O-acyl-2-lyso-sn-glycerol-3-phosphocholine
1-O-alkyl-2-lyso-sn-glycerol-3-phosphocholine + 1-O-acyl-2-acetyl-sn-glycerol-3-phosphocholine
show the reaction diagram
-
-
-
?
1-O-alkyl-2-acetyl-sn-glycerol-3-phosphocholine + H2O
1-O-alkyl-2-lyso-sn-glycerol-3-phosphocholine + acetate
show the reaction diagram
1-palmitoyl-2-(6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl)-sn-glycero-3-phosphocholine + cerebrosterol
1-palmitoyl-sn-glycero-3-phosphocholine + cerebrosteryl-(6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]hexanoyl)-ester
show the reaction diagram
-
i.e. 16:0-6:0 NBD-PC + (24S)-hydroxycholesterol
-
-
?
1-palmitoyl-2-20:4-sn-glycero-3-phosphocholine + cholesterol
?
show the reaction diagram
-
-
-
-
?
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine + cholesterol
1-palmitoyl-sn-glycero-3-phosphocholine + cholesteryl oleate
show the reaction diagram
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine + cholesterol
?
show the reaction diagram
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine + dehydroergosterol
1-palmitoyl-glycerophosphocholine + dehydroergosterol 3-O-oleoyl ester
show the reaction diagram
dehydroergosterol is a naturally occurring fluorescent sterol, that is esterified by enzyme LCAT, although at a slower rate than esterification of cholesterol, assay method development, overview
-
-
?
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoserine + cholesterol
?
show the reaction diagram
-
-
-
-
?
2-sn-phosphorylcholinediacylglycerol + cholesterol
?
show the reaction diagram
-
lower activity than the natural substrate
-
?
cholesterol + 1-O-hexadecyl-2-oleylphosphatidylcholine
cholesteryl oleate + 1-O-hexadecylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-palmitoyl-2-arachidonoylphosphatidylcholine
cholesteryl arachidonate + 1-palmitoylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-palmitoyl-2-docosahexaenoylphosphatidylcholine
cholesteryl docosahexaenoate + 1-palmitoylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-palmitoyl-2-eicosapentaenoylphosphatidylcholine
cholesteryl eicosapentaenoate + 1-palmitoylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-palmitoyl-2-linoleoylphosphatidylcholine
cholesteryl linoleate + 1-palmitoylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-palmitoyl-2-oleoylphosphatidylcholine
cholesteryl oleate + 1-palmitoylglycerophosphocholine
show the reaction diagram
cholesterol + 1-palmitoyl-2-phytanoylphosphatidylcholine
cholesteryl phytanoate + 1-palmitoylglycerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + 1-phytanoyl-2-palmitoylphosphatidylcholine
cholesteryl palmitate + 1-phytanylglyerophosphocholine
show the reaction diagram
-
-
-
?
cholesterol + egg lecithin
cholesteryl ester + ?
show the reaction diagram
-
-
-
-
?
dioleoyl-phosphatidyl choline + cholesterol
1-oleoyl-phosphatidyl choline + cholesteryl oleate
show the reaction diagram
-
-
-
-
?
HDL + cholesterol
?
show the reaction diagram
-
-
-
-
?
p-nitrophenol butyrate + H2O
p-nitrophenol + butyric acid
show the reaction diagram
-
esterase activity
-
?
phosphatidylcholine + 24-hydroxycholesterol
1-acylglycerophosphocholine + a 24-hydroxycholesterol 3-O-acyl ester
show the reaction diagram
-
-
-
-
?
phosphatidylcholine + a sterol
1-acylglycerophosphocholine + a sterol ester
show the reaction diagram
phosphatidylcholine + cerebrosterol
1-acylglycerophosphocholine + cerebrosteryl ester
show the reaction diagram
-
i.e. (24S)-hydroxycholesterol
-
-
?
phosphatidylcholine + cholesterol
1-acylglycerophosphocholine + cholesterol ester
show the reaction diagram
-
-
-
-
?
phosphatidylcholine + cholesterol
1-acylglycerophosphocholine + cholesteryl ester
show the reaction diagram
phosphatidylcholine + cholesterol
3-acylglycerophosphocholine + ?
show the reaction diagram
-
isozyme abnormality cause hepatosplenic schistosomiasis mansoni, important in lipoprotein metabolism and cholesterol transport, cholesterol is trapped in the HDL particles, enzyme transfers a long-chain fatty acyl residue from the sn-2 position of phosphatidylcholine, i.e. lecithin, to the 3-beta-hydroxyl group of cholesterol producing lysophosphatidylcholine or lysolecithin and cholesteryl ester, predominantly on HDL containing the activator apolipoprotein A-I
-
-
?
phosphatidylcholine + cholesterol
cholesteryl ester + lysophosphatidylcholine
show the reaction diagram
phosphatidylcholine + cholesterol
lysolecithin + cholesteryl ester
show the reaction diagram
-
enzyme transfers a long-chain fatty acyl residue from the sn-2 position of phosphatidylcholine, i.e. lecithin, to the 3-beta-hydroxyl group of cholesterol producing lysophosphatidylcholine or lysolecithin and cholesteryl ester, predominantly on HDL containing the activator apolipoprotein A-I
-
-
?
phosphatidylcholine + cholesterol
lysophosphatidylcholine + cholesteryl ester
show the reaction diagram
-
enzyme transfers a long-chain fatty acyl residue from the sn-2 position of phosphatidylcholine, i.e. lecithin, to the 3-beta-hydroxyl group of cholesterol producing lysophosphatidylcholine or lysolecithin and cholesteryl ester, predominantly on HDL containing the activator apolipoprotein A-I
-
-
?
phosphatidylcholine + sitosterol
1-acylglycerophosphocholine + sitosteryl ester
show the reaction diagram
-
purified recombinant enzyme
-
-
?
phosphatidylcholine + sterol
1-acylglycerophosphocholine + sterol ester
show the reaction diagram
phosphatidylethanolamine + cholesterol
cholesteryl ester + lysophosphatidylethanolamine
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
phosphatidylcholine + a sterol
1-acylglycerophosphocholine + a sterol ester
show the reaction diagram
phosphatidylcholine + cerebrosterol
1-acylglycerophosphocholine + cerebrosteryl ester
show the reaction diagram
-
i.e. (24S)-hydroxycholesterol
-
-
?
phosphatidylcholine + cholesterol
1-acylglycerophosphocholine + cholesterol ester
show the reaction diagram
-
-
-
-
?
phosphatidylcholine + cholesterol
1-acylglycerophosphocholine + cholesteryl ester
show the reaction diagram
phosphatidylcholine + cholesterol
3-acylglycerophosphocholine + ?
show the reaction diagram
-
isozyme abnormality cause hepatosplenic schistosomiasis mansoni, important in lipoprotein metabolism and cholesterol transport, cholesterol is trapped in the HDL particles, enzyme transfers a long-chain fatty acyl residue from the sn-2 position of phosphatidylcholine, i.e. lecithin, to the 3-beta-hydroxyl group of cholesterol producing lysophosphatidylcholine or lysolecithin and cholesteryl ester, predominantly on HDL containing the activator apolipoprotein A-I
-
-
?
phosphatidylcholine + cholesterol
cholesteryl ester + lysophosphatidylcholine
show the reaction diagram
phosphatidylcholine + sterol
1-acylglycerophosphocholine + sterol ester
show the reaction diagram
additional information
?
-
COFACTOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
apolipoprotein A1
-
-
-
additional information
-
hydrolysis of platelet-activating factor by LCAT does not require apolipoprotein as cofactor
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
Co2+
-
stimulates phospholipase reaction and cholesterol esterification, EDTA suppresses stimulation
Cu2+
-
stimulates phospholipase reaction and cholesterol esterification, EDTA suppresses stimulation
Ni2+
-
stimulates phospholipase reaction and cholesterol esterification, EDTA suppresses stimulation
Zn2+
-
stimulates phospholipase reaction and cholesterol esterification, EDTA suppresses stimulation
additional information
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1,2-diphytanoylphosphatidylcholine
-
competitive inhibitor
1-O-hexadecyl-2-O-oleoylphosphatidylcholine
-
competitive inhibitor
2-mercaptoethanol
-
at high concentration
4-(2-aminoethyl)benzenesulfonylfluoride
-
complete inhibition at 10 mM of the activity in plasma
5,5'-dithiobis(2-nitrobenzoic acid)
-
-
alpha2-Macroglobulin
-
association of this protein with LCAT inhibits the activity of LCAT, and may have a role in its catabolism
-
apolipoprotein A-II
-
-
-
apolipoprotein C-2
-
inhibitor in high concentration
-
carnitine
-
-
castanospermine
-
reduces the maximum reaction velocity but not the Km
ceramide phosphate
-
stronger physiologic inhibitory effect compared to sphingomyelin
chlorpromazine
-
-
cholesteryl ester transfer protein
-
56% inhibition of the activity in plasma
-
Cu2+
-
-
cysteine
-
-
deoxynojirimycin
-
reduces the maximum reaction velocity but not the Km
diisopropyl fluorophosphate
dithiothreitol
-
at high concentration
DTNB
-
83% inhibition at 1.4 mM of the activity in plasma
glycated discoidal (A-I)rHDL
-
after modification of lipid-free apoA-I arginine, lysine and tryptophan residues by glucose
-
H2O2
-
-
Haptoglobin
-
binds apolipoprotein A-I and E, via its Hpt beta-subunit, and impairs its stimulation of LCAT, mechanism, overview. The affinity of Hpt for ApoE is higher than that for ApoA-I. Hpt also impairs human hepatoblastoma-derived cell uptake of cholesterol from proteoliposomes containing ApoE or ApoA-I
-
lysolecithin
-
inhibition of LCAT activity
lysophosphatidylcholine
-
both enantiomers, inhibition reversed by albumin
methyldeoxynojirimycin
-
reduces the maximum reaction velocity but not the Km
p-hydroxymercuribenzoate
Phenylarsenoxide derivatives
-
-
phenylmethylsulfonyl fluoride
reduced glutathione
-
-
Serum albumin
-
inhibits LAT activity
-
Sn-2-Difluoroketone phosphatidylcholine analogues
-
IC50 values between 0.5 and 0.028 mM
-
sphingomyelin
Triton X-100
-
inhibition reversed by albumin
tunicamycin
-
reduces the maximum reaction velocity but not the Km
unesterified cholesterol-(apo lipoprotein A-I-)HDL
-
the unesterified cholesterol is deuterium-labeled
-
additional information
-
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
3-{[5-(ethylsulfanyl)-1,3,4-thiadiazol-2-yl]sulfanyl}pyrazine-2-carbonitrile
albumin
-
alpha-linolenic acid
-
a moderate dietary intake of myristic acid (1.8% total energy) and alpha-linolenic acid (0.9% total energy) increases LCAT activity
Apolipoprotein A-I
-
apolipoprotein A-IV
-
-
-
Apolipoprotein C-1
-
apolipoprotein E
-
atorvastatin
-
significant increase in serum LCAT activity in patients with hyperlipoproteinemia during atorvastatin therapy for 3 months, overview
ceramide
-
30% activation, stimulation of synthesis of 20:4 cholesteryl ester and 18:2 cholesteryl ester, but not of 16:0 cholesteryl ester, a phosphocholine ether matrix abolishes the activation effect
LCAT activating peptide LAP642
-
an amphiphilic peptide in place of apolipoprotein A-I is used as the lipid emulsifier and enzyme LCAT activator. The peptide forms stable complexes with phosphatidylcholine and sterol that react suitably well with enzyme LCAT, Km is 0.006 mM
-
low-density lipoprotein
-
required for acylation of lysolecithin
-
myristic acid
-
a moderate dietary intake of myristic acid (1.8% total energy) and alpha-linolenic acid (0.9% total energy) increases LCAT activity
Reducing agents
-
necessary for maximal activity
-
Serum albumin
-
activates LCAT activity
-
additional information
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.14 - 20.5
1,2-bis-(1-pyrenebutanoyl)-sn-glycero-3-phosphocholine
0.19
1,2-bis-(1-pyrenebutanoyl)-sn-gycero-3-phosphocholine
-
phospholipase activity of recombinant enzyme expressed in baby hamster kidney cells
-
0.00056
1,2-bis[4-(1-pyreno)butanoyl]-sn-glycero-3-phosphocholine
fluorescent assay, pH 7.4, 25C, recombinant enzyme
-
0.0006 - 0.027
1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine
0.000091
apo-A-1
-
-
-
0.57 - 21.4
cholesterol
0.003
dehydroergosterol
-
recombinant enzyme, pH 7.4, 37C
0.000026 - 0.0006
phosphatidylcholine
additional information
additional information
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
additional information
-
inhibition kinetics
-
IC50 VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.5 - 0.028
Sn-2-Difluoroketone phosphatidylcholine analogues
Homo sapiens
-
IC50 values between 0.5 and 0.028 mM
-
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.00078
purified recombinant enzyme, pH 7.5, 37C
0.001
-
recombinant enzyme
0.01
-
in cholesterol-lecithin vesicles activated with apolipoprotein A1, at 37C and pH 7.1
0.06
-
-
0.08
-
-
0.14
-
in cholesterol-lecithin vesicles activated with rat apolipoprotein A1
0.15
-
in cholesterol-lecithin vesicles activated with porcine apolipoprotein A1
0.217
-
purified recombinant enzyme, pH 7.4, 37C, substrates are 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and dehydroergosterol
0.27
-
in cholesterol-lecithin vesicles activated with apolipoprotein A1
0.34
-
at 37C in cholesterol-lecithin vesicles
0.402
-
purified recombinant enzyme, substrate cholesterol
0.42
-
purified recombinant enzyme, substrate sitosterol
0.43
-
recombinant C-terminal histidine tagged enzyme, similar to plasma enzyme
0.47
-
HDL substrate at 37C
0.54
-
at 37C and pH: 7.4
0.55
-
in cholesterol-lecithin vesicles activated with human apolipoprotein A1
0.56
-
in cholesterol-lecithin vesicles activated with human apolipoprotein A1
0.59
-
in cholesterol-lecithin vesicles activated with human apolipoprotein A1
0.867
-
purified recombinant enzyme, pH 7.4, 37C, substrates are 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine and cholesterol
2.7
-
in cholesterol-lecithin vesicles
3.47
-
in cholesterol-lecithin vesicles activated with apolipoprotein A1
53
-
in cholesterol-lecithin vesicles activated with apolipoprotein A1
137
-
in cholesterol-lecithin vesicles
additional information
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
6 - 9
-
activity is independent of pH in this range
7.5 - 8
-
lecithin-cholesterol acyltransferase activity
7.8
-
-
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
30
-
assay at
pI VALUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.9 - 4.8
-
plasma isozymes
4.51 - 5.26
-
isolectric focusing of 9 isozymes, the intensity of spot e (pI 4.86) is 3fold higher in amyotrophic lateral sclerosis compared to control
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
secretion to plasma
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
49000 - 62000
-
recombinant enzyme, gel filtration
52000
-
after cleavage of carbohydrate moiety
59000
-
sedimentation equilibrium ultracentrifugation
60000
-
sedimentation equilibrium ultracentrifugation
63000
-
sedimentation equilibrium
70000
-
gel filtration
83000
-
gradient gel electrophoresis
additional information
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
-
Trp61 functions as a component of the interfacial recognition domain of the enzyme, comparison of conformational changes upon substrate binding of wild-type enzyme and mutant T123I, overview
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
pH STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
3.5 - 7
-
partially purified enzyme is stable down to pH 3.5
486997
TEMPERATURE STABILITY
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37
-
0.4 mM phosphate buffer, pH 7.4, ionic strength 0.001: stable up to 6 h, 39 mM phosphate buffer, pH 7.4, ionic strength 0.1: 90% loss of activity
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
apo-A-1 prevents inactivation in phosphate buffer
-
apo-A-1, albumin and lecithin-cholesterol vesicles stabilize
-
complete loss of activity upon lyophilization
-
glycerol stabilizes during storage at -20C
-
inactivation at the air-water interface is prevented by buffered media of low ionic strength
-
lability to freezing and thawing and to high ionic strength
-
lecithin-cholesterol vesicles stabilize against inactivation to a lesser extent than apo-A-1
-
less stable than human enzyme to ionic strength
-
remarkably stable in buffers of very low ionic strength
-
the purified enzyme is instable
-
unaffected by repeated freezing and thawing
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-20C, 90% of activity is retained, 4 weeks
-
-80C, purified recombinant enzyme, 1-2 mg/ml protein in phosphate buffer with 10% glycerol, the enzyme retains full activity for at least 6 months and is stable to repeated freeze-thaw cycles
-
0C, 30 days
-
4C, 0.4 mM phosphate buffer, 4 mM 2-mercaptoethanol, under N2, 4 weeks
-
4C, 100 h
-
4C, 20% loss of activity, 24 h
-
4C, 60 days
-
4C, low ionic strength buffer, under nitrogen, 4 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
affinity chromatography using a p-aminophenylarsenoxide-linked phase
-
affinity chromatography using an anti-apolipoprotein D immunoglobulin-Sepharose column
-
antibody affinity chromatography
-
extraction from cell membranes with Tween 20, purification by Sephadex G-100 and DEAE-Sepharose columns
-
further purification of a recombinant enzyme using ACA-44 allows to remove a proteoglycan contaminant
-
HDL-Sepharose, agglutinin-Sepharose chromatography and polyacrylamide electrophoresis
-
native enzyme from plasma
-
native enzyme from plasma to homogeneity
-
partial, using phenyl-Sepharose chromatography
phenyl-Sepharose and hydroxyapatite column chromatography
-
precipitation from plasma and purification by phenyl-agarose, DEAE-cellulose and hydroxyapatite columns
-
precipitation from plasma and purification by phenyl-sepharose and concanavalin A-Sepharose columns
-
precipitation from plasma with dextran sulfate-Ca2+ followed by DEAE-Sephadex A-50 and ion-exchange chromatography, rat enzyme is not absorbed on hydroxyapatite, which is employed for the purification of the human enzyme
-
precipitation from plasma with dextran sulfate-Mg2+ followed by several chromatographic steps, including DEAE-Sepharose, phenyl-Sepharose and affinity chromatography
purification of a recombinant C-terminal histidine tagged enzyme using a cobalt affinity resin
-
purification of recombinant enzymes
-
recombinant enzyme from HEK-293Fcells
-
recombinant enzyme from human lung cell line H1299 culture medium
recombinant enzyme from Spodoptera frugiperda Sf21 cells by immunoaffinity chromatography, recombinant His-tagged peptide TgLCAT(106-202) from Escherichia coli strain M15 by nickel affinity chromatography
recombinant His-tagged wild-type and mutant enzymes from Escherichia coli strain BL21(DE3)pLysS by nickel affinity chromatography
-
recombinant LCAT from the conditioned medium of engineered human lung cells
-
recombinant partially deglycosylated His-tagged enzyme from HEK-293 GnTI cells by nickel affinity chromatography to homogeneity
-
recombinant wild-type and mutant enzymes from COS-1 cells
-
Sephadex G-100, hydroxyapatite and affinity column chromatography
-
several chromatographic steps, including affinity chromatography
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
expression in baby hamster kidney cells
-
expression in baby hamster kidney cells; expression in hepatic Mc-7777 cells
-
expression in Chinese hamster ovary cells, recombinant enzyme is structurally similar to plasma LCAT
-
expression in mice and rabbit
-
expression of a C-terminal histidine tagged enzyme in Chinese hamster ovary cells
-
expression of C-terminally His6-tagged LCAT in CHO cells
-
expression of mutant enzymes in baby hamster kidney cells
-
expression of mutant enzymes in COS-6 cells
-
expression of the naturally occurring mutants T123I and N228K in COS-1 and CHO cells
-
expression of truncated enzymes in COS-1 cells
-
expression of wild-type enzyme and N-terminally truncated enzymes in COS-7 cells, secretion to the cell culture
-
gene lcat, located on chromosome VIII, DNA and amino acid sequence determination and analyis, sequence comparisons and phylogenetic analysis, recombinant stable expression of HA- or YFP-tagged enzyme in Toxoplasma gondii, recombinant expression of His-tagged peptide TgLCAT(106-202) in Escherichia coli strain M15, recombinant expression in insect cells, using baculovirus transfection, and secretion to the culture medium
gene lcat, recombinant expression of His-tagged wild-type and mutant enzymes in Escherichia coli strain BL21(DE3)pLysS, recombinant expression of the enzyme in apoI-deficient mice, recombinant expression in CHO cells
-
gene lcat, stable recombinant expression in HEK-293Fcells
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gene lcat, use of baculovirus as a transducing vector to express the C-or N-terminally His-tagged enzyme in insect cell line Sf9, and to transiently express tagged LCAT in human HEK-293 GnTI cells, deficient in N-acetylglucosaminyl transferase I and expression of the recombinant protein as a high-mannose glycoform (of the MAN9GlcNAc2 or MAN5GlcNAc2 types) suitable for deglycosylation by Endo H, which leaves a single N-acetyl glucosamine (GlcNAc) residue at the glycosylation sites, to avoid protein aggregation coccuring with fully deglycosylated protein. Or expression in HEK-293 6E cells. TEV or Factor Xa sites are introduced for tag cleavage. The enzyme is secreted. Method development and evaluation, overview
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LCAT overexpression in transgenic mouse J774 macrophages
optimization of expression in human lung cell line H1299, enzyme is secreted to the culture medium
overexpression of LCAT in male nonhuman primate squirrel monkeys, Saimiri sciureus, using an adenoviral vector affects the lipoprotein metabolism, leading to an antiatherogenic lipoprotein profile by increasing HDL cholesterol and lowering ApoB, overview. Increased LCAT activity is also associated with a change in the size of HDL
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overexpression of LCAT in rabbit hepatocytes
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stable expression in CHO cells, expression in insect cells via baculovirus infection, glycosylation pattern of the recombinant enzymes differs from the wild-type, overview
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transient expression of wild-type and mutant enzymes in COS-1 cells
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EXPRESSION
ORGANISM
UNIPROT
LITERATURE
persons with normal LCAT alleles are also reported to experience reductions in LCAT activity in conjunction with certain diseases including coronary artery disease, diabetes, kidney disease, rheumatoid arthritis, and anemia
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ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
C31Y
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site-directed mutagenesis, the mutation renders the enzyme more stable and active than the native form
E149A
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site-directed mutagenesis, mutation alters the human enzyme residue to the corresponding residue of the rat sequence, 2.9fold increased cholesteryl ester formation activity, 5.5fold increased phospholipase A2 activity in the mutant compared to the wild-type enzyme
E149A/Y292H/W294F
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site-directed mutagenesis, mutation alters the human enzyme residues to the corresponding residues of the rat sequence, increased cholesteryl ester formation activity and phospholipase A2 activity with 1-palmitoyl-2-20:4-sn-glycero-3-phosphocholine, decreased activities with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine compared to the wild-type enzyme
T123I
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naturally occuring mutation involved in the fish eye disease, conformational changes upon substrate binding is altered in mutant T123I compared to the wild-type enzyme, overview
V309M
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naturally occuring mutation in exon 6, the rare enzyme genetic disorder, familial LCAT deficiency, leads to corneal opacities and proteinuria with renal failure, phenotype analysis of a Polish family, the patients show 10% of control enzyme activity and highly reduced enzyme concentrations, low total HDL-cholesterol and cholesteryl ester concentrations, decreased apo AI and apo AII serum levels, low LDL-cholesterol and apoB and Lp levels, and increased oleate/linoleate ratios, in cholestryl esters, phenotype, overview
Y292H/W294F
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site-directed mutagenesis, mutation alters the human enzyme residues to the corresponding residues of the rat sequence, 1.4fold increased cholesteryl ester formation activity, 2.8fold increased phospholipase A2 activity in the mutant compared to the wild-type enzyme
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
drug development
medicine
Show AA Sequence (121 entries)
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