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Literature summary for 3.1.4.54 extracted from

  • Wang, J.; Okamoto, Y.; Morishita, J.; Tsuboi, K.; Miyatake, A.; Ueda, N.
    Functional analysis of the purified anandamide-generating phospholipase D as a member of the metallo-beta-lactamase family (2006), J. Biol. Chem., 281, 12325-12335.
    View publication on PubMed
Search for more literature for 3.1.4.54

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
367stop complete loss of activity Rattus norvegicus
377stop complete loss of activity Rattus norvegicus
387stop Vmax is 20% of wild-type activity, slight decrease in Km-value for N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine Rattus norvegicus
C170S Vmax is 44% of wild-type activity Rattus norvegicus
C222S Vmax is 88% of wild-type activity Rattus norvegicus
C224S Vmax is 7.8% of wild-type activity Rattus norvegicus
C237S Vmax is 90% of wild-type activity Rattus norvegicus
C255S Vmax is 88% of wild-type activity Rattus norvegicus
C288S Vmax is 87% of wild-type activity Rattus norvegicus
D284N complete loss of activity Rattus norvegicus
D389N Vmax is about 87% of wild-type activity, single nucleotide polymorphism mutant Homo sapiens
DELTAN138 complete loss of activity Rattus norvegicus
DELTAN55 Vmax is 28% of wild-type activity, slight decrease in Km-value for N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine Rattus norvegicus
DELTAN85 complete loss of activity Rattus norvegicus
H185N complete loss of activity Rattus norvegicus
H187N complete loss of activity Rattus norvegicus
H189N complete loss of activity Rattus norvegicus
H190N complete loss of activity Rattus norvegicus
H253N complete loss of activity Rattus norvegicus
H321N complete loss of activity Rattus norvegicus
H331N specific activity is about 4% of wild-type value Rattus norvegicus
H343N specific activity is similar to wild-type value Rattus norvegicus
H353N specific activity is similar to wild-type value Rattus norvegicus
H380R complete loss of activity, single nucleotide polymorphism mutant Homo sapiens
H380R Vmax is about 4% of wild-type activity, single nucleotide polymorphism mutant Rattus norvegicus
L207F complete loss of activity, single nucleotide polymorphism mutant of Homo sapiens
L207F Vmax is about 1% of wild-type activity, single nucleotide polymorphism mutant Rattus norvegicus
S152A Vmax is about 84% of wild-type activity, single nucleotide polymorphism mutant Homo sapiens

General Stability

General Stability Organism
1% (w/v) octyl glucoside improves the stability of the purified enzyme Rattus norvegicus
one cycle of freezing and thawing causes loss of the enzyme activity up to 50% Rattus norvegicus

Inhibitors

Inhibitors Comment Organism Structure
Cu2+
-
 Rattus norvegicus
Fe2+
-
 Rattus norvegicus
Hg2+
-
 Rattus norvegicus
Zn2+
-
 Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0029
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme D389N Homo sapiens
0.0029
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme DELTAN55 Rattus norvegicus
0.0036
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme 387stop Rattus norvegicus
0.0037
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme S152A Homo sapiens
0.0042
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C237S Rattus norvegicus
0.0043
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C122S Rattus norvegicus
0.0045
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C288S Rattus norvegicus
0.0046
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, wild-type enzyme Homo sapiens
0.0047
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, wild-type enzyme Rattus norvegicus
0.0047
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C170S Rattus norvegicus
0.0061
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C255S Rattus norvegicus
0.0142
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme H380R Rattus norvegicus
0.0143
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C224S Rattus norvegicus
0.0647
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme L207F Rattus norvegicus

Metals/Ions

Metals/Ions Comment Organism Structure
Ba2+ stimulates Rattus norvegicus
Ca2+ stimulates at millimolar concentrations, EC50: 1.4 mM Rattus norvegicus
Co2+ stimulates Rattus norvegicus
Mg2+ stimulates at millimolar concentrations, EC50: 2.3 mM Rattus norvegicus
Mn2+ stimulates Rattus norvegicus
Sr2+ stimulates Rattus norvegicus
Zn2+ the enzyme belongs to the metallo-beta-lactamase family, presence of catalytically important zinc Rattus norvegicus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
46000
-
 x * 46000, SDS-PAGE Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Homo sapiens Q6IQ20
-
-
Rattus norvegicus Q769K2
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Rattus norvegicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.6
-
 37°C, pH 7.4, substrate: N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine, wild-type enzyme Rattus norvegicus

Storage Stability

Storage Stability Organism
-80°C, in the presence of 1% octyl glucoside Rattus norvegicus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
additional information low activity with N-acetylphosphatidylethanolamine or N-formylphosphatidylethanolamine. The specific activities with N-palmitoyl-lyso-phosphatidylethanolamine and glycerophospho(N-palmitoyl)ethanolamine are 4% and 1%, respectively, of that with N-palmitoylphosphatidylethanolamine. Furthermore, N-palmitoylethanolamine phosphate is totally inactive. N-Palmitoyl-phosphatidylserine, phosphatidylethanol, and phosphatidylbutanol are hydrolyzed at less than 0.4% of the activity as compared with N-palmitoylphosphatidylethanolamine Rattus norvegicus ?
-
 ?
N-arachidonoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-arachidonoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-butanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-butanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
N-decanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-decanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
N-hexanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-hexanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
N-lauroyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-lauroylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-myristoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-myristoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-octanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-octanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
N-oleoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-oleoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Homo sapiens N-palmitoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-palmitoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
N-stearoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-stearoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?

Subunits

Subunits Comment Organism
? x * 46000, SDS-PAGE Rattus norvegicus

Synonyms

Synonyms Comment Organism
anandamide-generating phospholipase D
-
 Rattus norvegicus