Information on EC 3.1.2.22 - palmitoyl[protein] hydrolase

New: Word Map on EC 3.1.2.22
Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Specify your search results
Mark a special word or phrase in this record:
Search Reference ID:
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: Opisthokonta

EC NUMBER
COMMENTARY hide
3.1.2.22
-
RECOMMENDED NAME
GeneOntology No.
palmitoyl[protein] hydrolase
REACTION
REACTION DIAGRAM
COMMENTARY hide
ORGANISM
UNIPROT
LITERATURE
palmitoyl[protein] + H2O = palmitate + protein
show the reaction diagram
REACTION TYPE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
hydrolysis of thioester
PATHWAY
BRENDA Link
KEGG Link
MetaCyc Link
Fatty acid elongation
-
-
Metabolic pathways
-
-
SYSTEMATIC NAME
IUBMB Comments
palmitoyl[protein] hydrolase
Specific for long-chain thioesters of fatty acids. Hydrolyses fatty acids from S-acylated cysteine residues in proteins, palmitoyl cysteine and palmitoyl-CoA.
CAS REGISTRY NUMBER
COMMENTARY hide
150605-49-5
-
158210-90-3
hydrolase, acyl protein thioester (rat clone pRatPPT-44 precursor reduced) /palmitoyl-protein thioesterase (rat clone pRatPPT-44 precursor)
158210-92-5
hydrolase, acyl protein thioester (cattle clone pBovPPT-18 precursor reduced) /hydrolase, acyl protein thioester (ox clone pBovPPT-18 precursor reduced) /palmitoyl-protein thioesterase (bovine clone pBovPPT-17 precursor)
158210-93-6
hydrolase, acyl protein thioester (cattle pBovPPT-25 precursor reduced) /hydrolase, acyl protein thioester (ox pBovPPT-25 precursor reduced) /palmitoyl-protein thioesterase (bovine clone pBovPPT-25 precursor)
179801-10-6
hydrolase, acyl protein thioester (human clone pHuPPT-5') /GenBank L42809-derived protein GI 1160967 /palmitoyl-protein thioesterase (human clone pHuPPT-5')
199619-79-9
hydrolase, acyl protein thioester (human clone B lysosome-associated isoenzyme 2) /genBank AF020543-derived protein GI 2501961 /palmitoyl-protein thioesterase PPT2 (human clone B lysosome associated)
ORGANISM
COMMENTARY hide
LITERATURE
UNIPROT
SEQUENCE DB
SOURCE
-
-
-
Manually annotated by BRENDA team
strain deleted for ppt-1, potentially provides a model system for the study of human infantile neuronal ceroid lipofuscinosis
-
-
Manually annotated by BRENDA team
Miniature Dachshund
UniProt
Manually annotated by BRENDA team
cosmid; isozyme PPT1. gene pdf1
UniProt
Manually annotated by BRENDA team
GENERAL INFORMATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
malfunction
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
1-palmitoyl lysophosphatidylcholine + H2O
glycerophosphocholine + palmitate
show the reaction diagram
-
-
-
?
4-methylumbelliferyl 6-deoxy-6-thiopalmitoyl-beta-D-glucopyranoside + H2O
4-methylumbelliferyl 6-deoxy-6-thio-beta-D-glucopyranoside + palmitate
show the reaction diagram
-
-
-
?
4-methylumbelliferyl-6-S-palmitoyl-beta-D-glucoside + H2O
4-methylumbelliferol + palmitate + beta-D-glucose
show the reaction diagram
-
-
-
?
4-methylumbelliferyl-6-thiopalmitoyl-beta-D-glucoside + H2O
4-methylumbelliferol + palmitate + beta-D-glucose
show the reaction diagram
4-methylumbelliferyl-6-thiopalmitoyl-beta-D-glucoside + H2O
?
show the reaction diagram
Myristoyl-CoA + H2O
Myristate + CoA
show the reaction diagram
-
-
-
-
-
Palmitoyl-(alpha-subunit of the heterotrimeric G protein) + H2O
?
show the reaction diagram
-
-
-
-
-
Palmitoyl-CoA + H2O
Palmitate + CoA
show the reaction diagram
Palmitoyl-H-Ras + H2O
Palmitate + Ha-Ras
show the reaction diagram
palmitoyl-peptide + H2O
palmitate + peptide
show the reaction diagram
residue 3 to 14 of RGS4 protein with palmitoyl group attached at Cys12 of RGS4 protein
-
-
?
palmitoyl-protein + H2O
palmitate + protein
show the reaction diagram
palmitoyl-[protein] + H2O
palmitate + protein
show the reaction diagram
palmitoylthio-beta-glucoside + H2O
palmitate + thio-beta-glucoside
show the reaction diagram
-
-
-
?
S-palmitoyl-N-acetyl-O-carboxymethyl-cysteine + H2O
N-acetyl-O-carboxymethyl-cysteine + palmitate
show the reaction diagram
-
enzyme form PPT1, not PPT2
-
?
S-palmitoyl-N-acetylcysteamine + H2O
N-acetylcysteamine + palmitate
show the reaction diagram
-
enzyme form PPT1, not PPT2
-
?
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
palmitoyl-protein + H2O
palmitate + protein
show the reaction diagram
additional information
?
-
METALS and IONS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
additional information
activity independent of Ca2+ and Mg2+
INHIBITORS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
1-palmitoyl lysophosphatidylcholine
competitive inhibitor for acylprotein thioesterase activity
Chloroquine
-
inhibits the hydrolysis and leads to accumulation of lipid cysteine thioesters in the lysosomes of treated cells
Didemnin B
-
-
diethyl dicarbonate
-
-
dithiothreitol
-
-
KCl
-
-
Leupeptin
-
inhibits the hydrolysis and leads to accumulation of lipid cysteine thioesters in the lysosomes of treated cells but has no direct inhibitory effect on the enzyme
NaCl
-
-
palmitoyl-peptide
competitive inhibitor for lysophospholipase activity
Sodium vanadate
pdf1 mutant
additional information
ACTIVATING COMPOUND
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
additional information
-
Drosophila C-terminal Src kinase is an enhancer of the Ppt1 phenotype. Mutations in the fear of intimacy gene are also identified as enhancers. Mutations in zipper or mutations at the Drosophila IGF-II mRNA-binding protein locus enhance the degenerative phenotype. The tsg alleles tsg2 and tsg4 both enhance the rough eye phenotype produced by Ppt1 expression
-
KM VALUE [mM]
SUBSTRATE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0273
1-palmitoyl lysophosphatidylcholine
-
0.091
myristoyl-CoA
-
-
0.227
palmitoyl-beta-D-thioglucoside
-
-
0.017 - 0.039
palmitoyl-CoA
0.01 - 0.02
palmitoyl-H-Ras
-
-
-
0.00349
palmitoyl-peptide
-
Ki VALUE [mM]
INHIBITOR
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
IMAGE
0.0222
1-palmitoyl lysophosphatidylcholine
for acylprotein thioesterase activity
0.0323
palmitoyl-peptide
for lysophospholipase activity
SPECIFIC ACTIVITY [µmol/min/mg]
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
0.0055
substrate 4-methylumbelliferyl 6-deoxy-6-thiopalmitoyl-beta-D-glucopyranoside, cerebral cortex gray matter lysate, pH not specified in the publication, temperature not specified in the publication
0.067
-
-
1.62
substrate 1-palmitoyl lysophosphatidylcholine
1.72
-
whole cell, wild-type enzyme, substrate palmitoyl-CoA
7.49
-
whole cell, wild-type enzyme, substrate S-palmitoyl-beta-glucoside
27.3
substrate palmitoyl-peptide
additional information
-
-
pH OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
4
-
assay at
5.2
-
assay at
6.5 - 7
-
-
7
-
palmitoyl-H-Ras
TEMPERATURE OPTIMUM
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
22
-
assay at, room temperature
SOURCE TISSUE
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
SOURCE
-
transformed by Epstein-Barr virus
Manually annotated by BRENDA team
-
exclusively localized to the basolateral side of the polarized cells
Manually annotated by BRENDA team
-
normal fibroblasts and infantile neuronal ceroid lipofuscinosis fibroblasts, deficient of PPT1
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
enzyme expression slowly decreases during maturation
Manually annotated by BRENDA team
-
-
Manually annotated by BRENDA team
-
primary cortical neuron culture from tissue of PPT1 knockout mice that mimic infantile neuronal ceroid lipofuscinosis
Manually annotated by BRENDA team
additional information
LOCALIZATION
ORGANISM
UNIPROT
COMMENTARY hide
GeneOntology No.
LITERATURE
SOURCE
-
preferably targeted to in mature neurons
Manually annotated by BRENDA team
-
in contrast to the wild type enzyme, the mutant polypeptides are retained in the endoplasmic reticulum, most probably due to the misfolding of the mutant polypeptide
Manually annotated by BRENDA team
-
the most common mutation results in intracellular accumulation of the enzyme polypeptide and undetectable activity in the brain
Manually annotated by BRENDA team
-
cortical, in synaptic junctions of epileptic rats, nearly not in non-epileptic ones
Manually annotated by BRENDA team
additional information
MOLECULAR WEIGHT
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
37000
-
gel filtration
SUBUNITS
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
monomer
-
1 * 37000, SDS-PAGE
additional information
PPT1 forms oligomers
POSTTRANSLATIONAL MODIFICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
glycoprotein
proteolytic modification
enzyme contains 2 putative cleavage sites for the kex-related endopeptidase Krp1p, the precursor is proteolytically processed to form distinct Ppt1p and Dolpp1p domains, Arg354 is crucial for the processing
Crystallization/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
purified recombinant enzyme with and without bound palmitate, sitting drop vapor diffusion method, 12 mg/ml enzyme in 20 mM HEPES, pH 7.0, plus 150 mM NaCl, plus reservoir solution, containing 55% PEG 400, 100 mM Bis-Tris, pH 6.5, in a ratio 3:2, equilibrated against a250fold excess, X-ray multiwave length anomalous diffraction phasing for structure determinationand analysis, 2.25 A resolution
-
purified recombinant PPT2 15 mg/ml, sitting drop vapour diffusion method, 4°C, with equal volume of precipitant solution conataining 2 M ammonium sulfate, 100 mM sodium cacodylate, pH 5.5-6.5, 8% methyl pentanediol, X-ray diffraction structure determination and analysis at 2.7 A resolution
-
GENERAL STABILITY
ORGANISM
UNIPROT
LITERATURE
repeated freeze-thawing decreases activity
-
STORAGE STABILITY
ORGANISM
UNIPROT
LITERATURE
-80°C, 50 mM Tris/HCl, pH 7.4, more than one year, stable
4°C, stable for at least 3 weeks
-
Purification/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
by centrifugation, dialysis and gel filtration
-
GST affinity chromatography, GST-tag cleaved from GST-fusion protein
recombinant PPT2 from overexpressing insect cells, recombinant PPT2 mutant from COS cells
-
Cloned/COMMENTARY
ORGANISM
UNIPROT
LITERATURE
baculovirally expressed
-
construction of a UAS:GFP-Ppt1 transgenic fly line. The pUAST:GFP-Ppt1 vector overexpressed using Act5C-Gal4
-
DNA sequence determination and analysis, enzyme is encoded in frame with a second gene coding for an indispensable dolichol pyrophosphatase, gene pdf1, strains bearing a deletion of the entire pdf1 open reading frame are nonviable, due to Dolpp1p deficiency, and are rescued by pdf1 expression plasmid, mutations only in the palmitoyl protein thioesterase 1 domain results in cells abnormally sensitive to sodium vanadate and elevated extracellular pH, the pdf1-deficient phenotype is complemented by the human PPT1 gene
expressed in COS-1 cells
-
expression as glutathione S-transferase fusion protein in BHK and CHO cells, localization of the recombinant enzyme out side the lysosomes and endosomes
-
expression in endothelial cells
-
expression of enzyme carrying naturally occurring mutations in COS-1 cells
-
expression of the enzyme in Caco-2 cells, orientation to the basolateral side, also after endocytosis of the recombinant enzyme into Caco-2 cells
-
expression of wild-type and enzymes with naturally occurring mutations in COS cells as His-tagged enzyme, and in Sf9 cells, the latter via baculovirus infection
-
expression of wild-type and mutant in COS-1 cells
-
fragment corresponding to the entire coding region of PPT1 amplified from pCMV5-hPPT1, and cloned into XhoI sites in the polylinker region of expression vector pMSXND1, overexpressed in CHO cells. PPT1 injected intravenously into PPT1-deficient mice
-
gene ppt, DNA sequence determnation and analysis, identification of 8 natural mutations of different origins leading to enzyme deficiency
-
GST-fusion protein expressed in Escherichia coli BL21, expressed in HEK-293 cell
overexpressed in Drosophila melanogaster
-
overexpression of PPT2 in an insect-baculovirus expression system, secretion of the recombinant enzyme to the culture medium, expression of mutant PPT2 in COS cells, subcloning in Escherichia coli XL1-blue
-
PPT1-deficient fibroblasts stably transfected with cDNA encoding PPT1
-
PPT1-KO mice are generated by gene targeting in ES cells
-
EXPRESSION
ORGANISM
UNIPROT
LITERATURE
in rats fed a high-fat diet, palmitoyl-protein thioesterase PPT1 is upregulated. In a Sertoli-cell line cultured in a high-fat supplemented medium, PPT1 abundance Is accompanied by increases in the endocytic vesicle-associated protein, clathrin, and decreases in the tight junctional proteins, ZO-1 and occludin
ENGINEERING
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
A179T
-
loss-of-function allele, although endocytosis and endo-lysosomal trafficking does occur in Ppt1 mutant garland cells, there is a reduced level of uptake and a decreased rate of trafficking to the lysosomes
S77F
-
loss-of-function allele, although endocytosis and endo-lysosomal trafficking does occur in Ppt1 mutant garland cells, there is a reduced level of uptake and a decreased rate of trafficking to the lysosomes
D233N
-
site-directed mutagenesis, inactive mutant
E184K
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with the infantile form of neuronal ceroid lipofuscinosis, no binding of mannose 6-phosphate receptor
F84del
-
natural deletion mutation in gene ppt1, leading to enzyme deficiency and infantile progressive neurological defects
G118D
-
natural mutation in gene ppt1, leading to enzyme deficiency and infantile progressive neurological defects
G250V
-
naturally occurring mutation, recombinant enzyme shows 5.9% of the wild-type activity, mutation is associated with the juvenile form of neuronal ceroid lipofuscinosis, no binding of mannose 6-phosphate receptor
G42E
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with the infantile form of neuronal ceroid lipofuscinosis, no binding of mannose 6-phosphate receptor
H289A
-
site-directed mutagenesis, inactive mutant
H39Q
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with the infantile form of neuronal ceroid lipofuscinosis, no binding of mannose 6-phosphate receptor
L219Q
-
naturally occurring mutation, mutation is associated with the late onset form of infantile neuronal ceroid lipofuscinosis, the mutant enzymes shows minor altered intracellular localization in transfected cells and are localized in the presynaptic space and neuronal shaft, about 10fold reduced activity
N197Q
-
site-directed mutagenesis, mutation of a glycosylation site, slightly reduced activity
N197Q/N212Q
-
site-directed mutagenesis, mutation of glycosylation sites, slightly reduced activity, mutant shows 10% of the wild-type expression level
N197Q/N212Q/N232Q
-
site-directed mutagenesis, mutation of all glycosylation sites, inactive mutant
N197Q/N232Q
-
site-directed mutagenesis, mutation of glycosylation sites, reduced activity
N212Q
-
site-directed mutagenesis, mutation of a glycosylation site, slightly reduced activity
N212Q/N232Q
-
site-directed mutagenesis, mutation of glycosylation sites, highly reduced activity
N232Q
-
site-directed mutagenesis, mutation of a glycosylation site, slightly reduced activity
Q291X
-
natural mutation in gene ppt1, leading to enzyme deficiency and infantile progressive neurological defects
R122W
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with the infantile form of neuronal ceroid lipofuscinosis, no binding of mannose 6-phosphate receptor, accumulation of the recombinant mutant enzyme in the endoplamic reticulum due to decreased degradation turnover
R164X
-
natural mutation in gene ppt1, leading to enzyme deficiency and infantile progressive neurological defects
S115A
-
site-directed mutagenesis, inactive mutant
S119A
part of the catalytic triad, no activity
S199A
mutation shows 1-5% of normal activity
S199A/S214A
mutant enzyme with very low enzyme activity
S214A
mutant shows 31% of normal activity
S234A
mutation shows 1-5% of normal activity. Partial localization to lysosomes, although a major proportion is retained in the endoplasmic reticulum
Y109D
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with an unclassified form of neuronal ceroid lipofuscinosis
Y247H
-
naturally occurring mutation, recombinant enzyme is nearly inactive showing an activity below 2% of the wild-type activity, mutation is associated with an unclassified form of neuronal ceroid lipofuscinosis
additional information
APPLICATION
ORGANISM
UNIPROT
COMMENTARY hide
LITERATURE
medicine
nutrition
the litter sizes for females mated to obese males are significantly lower as compared to females mated with normal-diet-fed controls. Their serum high-density lipoprotein, low-density lipoprotein, cholesterol, and estradiol levels increase in obese males, but testosterone and follicle-stimulating hormone levels decrease. Testicular morphology disruptions include Sertoli-cell atrophy, disrupted tight junctions, and mitochondrial degeneration in spermatogenic cells. In rats fed a high-fat diet, palmitoyl-protein thioesterase PPT1 is upregulated. In a Sertoli-cell line cultured in a high-fat supplemented medium, PPT1 abundance Is accompanied by increases in the endocytic vesicle-associated protein, clathrin, and decreases in the tight junctional proteins, ZO-1 and occludin