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Literature summary 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

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.4.4 expression in Escherichia coli Rattus norvegicus
3.1.4.54 expression in Escherichia coli Rattus norvegicus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.4.4 367stop complete loss of activity Rattus norvegicus
3.1.4.4 377stop complete loss of activity Rattus norvegicus
3.1.4.4 387stop activity comparable to wild-type Rattus norvegicus
3.1.4.4 C170S activity comparable to wild-type Rattus norvegicus
3.1.4.4 C222S activity comparable to wild-type Rattus norvegicus
3.1.4.4 C224S considerable reduction in activity Rattus norvegicus
3.1.4.4 C237S activity comparable to wild-type Rattus norvegicus
3.1.4.4 C255S activity comparable to wild-type Rattus norvegicus
3.1.4.4 C288S activity comparable to wild-type Rattus norvegicus
3.1.4.4 D147N complete loss of activity Rattus norvegicus
3.1.4.4 D189N less than 0.1% of wild-type activity Rattus norvegicus
3.1.4.4 D284N complete loss of activity Rattus norvegicus
3.1.4.4 D389N single nucleotide polymorphism, activity comparable to wild-type Homo sapiens
3.1.4.4 DELTAN138 complete loss of activity Rattus norvegicus
3.1.4.4 DELTAN55 activity comparable to wild-type Rattus norvegicus
3.1.4.4 DELTAN85 complete loss of activity Rattus norvegicus
3.1.4.4 H185N less than 0.1% of wild-type activity Rattus norvegicus
3.1.4.4 H187N less than 0.1% of wild-type activity Rattus norvegicus
3.1.4.4 H190N less than 0.1% of wild-type activity Rattus norvegicus
3.1.4.4 H253N complete loss of activity Rattus norvegicus
3.1.4.4 H321N complete loss of activity Rattus norvegicus
3.1.4.4 H331N 4% of wild-type activity Rattus norvegicus
3.1.4.4 H343N activity comparable to wild-type Rattus norvegicus
3.1.4.4 H353N activity comparable to wild-type Rattus norvegicus
3.1.4.4 H380R single nucleotide polymorphism, complete loss of activity Homo sapiens
3.1.4.4 H380R single nucleotide polymorphism, low catalytic activity Rattus norvegicus
3.1.4.4 L207F single nucleotide polymorphism, complete loss of activity Homo sapiens
3.1.4.4 L207F single nucleotide polymorphism, low catalytic activity Rattus norvegicus
3.1.4.4 S152A single nucleotide polymorphism, activity comparable to wild-type Homo sapiens
3.1.4.54 367stop complete loss of activity Rattus norvegicus
3.1.4.54 377stop complete loss of activity Rattus norvegicus
3.1.4.54 387stop Vmax is 20% of wild-type activity, slight decrease in Km-value for N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine Rattus norvegicus
3.1.4.54 C170S Vmax is 44% of wild-type activity Rattus norvegicus
3.1.4.54 C222S Vmax is 88% of wild-type activity Rattus norvegicus
3.1.4.54 C224S Vmax is 7.8% of wild-type activity Rattus norvegicus
3.1.4.54 C237S Vmax is 90% of wild-type activity Rattus norvegicus
3.1.4.54 C255S Vmax is 88% of wild-type activity Rattus norvegicus
3.1.4.54 C288S Vmax is 87% of wild-type activity Rattus norvegicus
3.1.4.54 D284N complete loss of activity Rattus norvegicus
3.1.4.54 D389N Vmax is about 87% of wild-type activity, single nucleotide polymorphism mutant Homo sapiens
3.1.4.54 DELTAN138 complete loss of activity Rattus norvegicus
3.1.4.54 DELTAN55 Vmax is 28% of wild-type activity, slight decrease in Km-value for N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine Rattus norvegicus
3.1.4.54 DELTAN85 complete loss of activity Rattus norvegicus
3.1.4.54 H185N complete loss of activity Rattus norvegicus
3.1.4.54 H187N complete loss of activity Rattus norvegicus
3.1.4.54 H189N complete loss of activity Rattus norvegicus
3.1.4.54 H190N complete loss of activity Rattus norvegicus
3.1.4.54 H253N complete loss of activity Rattus norvegicus
3.1.4.54 H321N complete loss of activity Rattus norvegicus
3.1.4.54 H331N specific activity is about 4% of wild-type value Rattus norvegicus
3.1.4.54 H343N specific activity is similar to wild-type value Rattus norvegicus
3.1.4.54 H353N specific activity is similar to wild-type value Rattus norvegicus
3.1.4.54 H380R complete loss of activity, single nucleotide polymorphism mutant Homo sapiens
3.1.4.54 H380R Vmax is about 4% of wild-type activity, single nucleotide polymorphism mutant Rattus norvegicus
3.1.4.54 L207F complete loss of activity, single nucleotide polymorphism mutant of Homo sapiens
3.1.4.54 L207F Vmax is about 1% of wild-type activity, single nucleotide polymorphism mutant Rattus norvegicus
3.1.4.54 S152A Vmax is about 84% of wild-type activity, single nucleotide polymorphism mutant Homo sapiens

General Stability

EC Number General Stability Organism
3.1.4.54 1% (w/v) octyl glucoside improves the stability of the purified enzyme Rattus norvegicus
3.1.4.54 one cycle of freezing and thawing causes loss of the enzyme activity up to 50% Rattus norvegicus

Inhibitors

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

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.4.4 0.0029
-
 N-palmitoyl-phosphatidylethanolamine mutant D389N, 37°C, pH 7.4, presence of 10 mM MgCl2 Homo sapiens
3.1.4.4 0.0029
-
 N-palmitoyl-phosphatidylethanolamine mutant DELTAN55, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0036
-
 N-palmitoyl-phosphatidylethanolamine mutant 387stop, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0037
-
 N-palmitoyl-phosphatidylethanolamine mutant S152A, 37°C, pH 7.4, presence of 10 mM MgCl2 Homo sapiens
3.1.4.4 0.0042
-
 N-palmitoyl-phosphatidylethanolamine mutant C237S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0043
-
 N-palmitoyl-phosphatidylethanolamine mutant C222S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0045
-
 N-palmitoyl-phosphatidylethanolamine mutant C288S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0046
-
 N-palmitoyl-phosphatidylethanolamine wild-type, 37°C, pH 7.4, presence of 10 mM MgCl2 Homo sapiens
3.1.4.4 0.0047
-
 N-palmitoyl-phosphatidylethanolamine mutant C170S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0047
-
 N-palmitoyl-phosphatidylethanolamine wild-type, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0059
-
 N-palmitoyl-phosphatidylethanolamine wild-type, 37°C, pH 7.4 Rattus norvegicus
3.1.4.4 0.0061
-
 N-palmitoyl-phosphatidylethanolamine mutant C255S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0143
-
 N-palmitoyl-phosphatidylethanolamine mutant C224S, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.4 0.0211
-
 N-palmitoyl-phosphatidylethanolamine mutant H331N, 37°C, pH 7.4, presence of 10 mM MgCl2 Rattus norvegicus
3.1.4.54 0.0029
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme D389N Homo sapiens
3.1.4.54 0.0029
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme DELTAN55 Rattus norvegicus
3.1.4.54 0.0036
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme 387stop Rattus norvegicus
3.1.4.54 0.0037
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme S152A Homo sapiens
3.1.4.54 0.0042
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C237S Rattus norvegicus
3.1.4.54 0.0043
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C122S Rattus norvegicus
3.1.4.54 0.0045
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C288S Rattus norvegicus
3.1.4.54 0.0046
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, wild-type enzyme Homo sapiens
3.1.4.54 0.0047
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, wild-type enzyme Rattus norvegicus
3.1.4.54 0.0047
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C170S Rattus norvegicus
3.1.4.54 0.0061
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C255S Rattus norvegicus
3.1.4.54 0.0142
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme H380R Rattus norvegicus
3.1.4.54 0.0143
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme C224S Rattus norvegicus
3.1.4.54 0.0647
-
 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine 37°C, pH 7.4, mutant enzyme L207F Rattus norvegicus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.4.4 Ca2+ 50% activation at 2.3 mM Rattus norvegicus
3.1.4.4 Mg2+ 50% activation at 2.3 mM Rattus norvegicus
3.1.4.4 Zn2+ catalytically active Zn2+ Rattus norvegicus
3.1.4.54 Ba2+ stimulates Rattus norvegicus
3.1.4.54 Ca2+ stimulates at millimolar concentrations, EC50: 1.4 mM Rattus norvegicus
3.1.4.54 Co2+ stimulates Rattus norvegicus
3.1.4.54 Mg2+ stimulates at millimolar concentrations, EC50: 2.3 mM Rattus norvegicus
3.1.4.54 Mn2+ stimulates Rattus norvegicus
3.1.4.54 Sr2+ stimulates Rattus norvegicus
3.1.4.54 Zn2+ the enzyme belongs to the metallo-beta-lactamase family, presence of catalytically important zinc Rattus norvegicus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.4.4 46000
-
 x * 46000, SDS-PAGE Rattus norvegicus
3.1.4.54 46000
-
 x * 46000, SDS-PAGE Rattus norvegicus

Organism

EC Number Organism UniProt Comment Textmining
3.1.4.4 Homo sapiens
-
 N-acylethanolamine-specific phospholipase D, NAPE-PLD
-
3.1.4.4 Rattus norvegicus
-
 N-acylethanolamine-specific phospholipase D, NAPE-PLD
-
3.1.4.54 Homo sapiens Q6IQ20
-
-
3.1.4.54 Rattus norvegicus Q769K2
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.1.4.4 recombinant enzyme Rattus norvegicus
3.1.4.54
-
 Rattus norvegicus

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.1.4.4 2.016
-
 37°C, pH 7.4, presence of 10 mM MgCl2, substrate N-palmitoyl-phosphatidylethanolamine Rattus norvegicus
3.1.4.4 2.4
-
 37°C, pH 7.4, presence of 10 mM MgCl2, substrate N-lauroyl-phosphatidylethanolamine Rattus norvegicus
3.1.4.54 2.6
-
 37°C, pH 7.4, substrate: N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine, wild-type enzyme Rattus norvegicus

Storage Stability

EC Number Storage Stability Organism
3.1.4.4 unstable, one cycle of freezeing and thawing causes loss of up to 50% of enzymic activity. Addition of 1% w/v octyl glucoside improves stability Homo sapiens
3.1.4.4 unstable, one cycle of freezing and thawing causes loss of up to 50% of enzymic activity. Addition of 1% w/v octyl glucoside improves stability Rattus norvegicus
3.1.4.54 -80°C, in the presence of 1% octyl glucoside Rattus norvegicus

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.4.4 glycerophospho-(N-palmitoyl)ethanolamine + H2O
-
 Rattus norvegicus N-palmitoylethanolamine + ? 1% of the activity with N-palmitoyl-phosphatidylethanolamine ?
3.1.4.4 additional information no substrate: N-palmitoylethanolamine phosphate, phosphatidylcholine, phosphatidylserine, phosphatidylinositol Rattus norvegicus ?
-
 ?
3.1.4.4 N-acyl-phosphatidylethanolamine + H2O high specificity for N-acyl-phosphatidylethanolamines without selectivity for long chain or medium chain N-acyl species Homo sapiens N-acylethanolamine + phosphatidate
-
 ?
3.1.4.4 N-acyl-phosphatidylethanolamine + H2O high specificity for N-acyl-phosphatidylethanolamines without selectivity for long chain or medium chain N-acyl species Rattus norvegicus N-acylethanolamine + phosphatidate
-
 ?
3.1.4.4 N-lauroyl-phosphatidylethanolamine + H2O
-
 Homo sapiens phosphatidic acid + N-lauroylethanolamine
-
 ?
3.1.4.4 N-lauroyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus phosphatidic acid + N-lauroylethanolamine
-
 ?
3.1.4.4 N-palmitoyl-lyso-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-palmitoylethanolamine + sn-glycerol 3-phosphate 4% of the activity with N-palmitoyl-phosphatidylethanolamine ?
3.1.4.4 N-palmitoyl-phosphatidylethanolamine + H2O
-
 Homo sapiens phosphatidic acid + N-palmitoylethanolamine
-
 ?
3.1.4.4 N-palmitoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus phosphatidic acid + N-palmitoylethanolamine
-
 ?
3.1.4.54 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 ?
-
 ?
3.1.4.54 N-arachidonoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-arachidonoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-butanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-butanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-decanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-decanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-hexanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-hexanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-lauroyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-lauroylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-myristoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-myristoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-octanoyl-1-palmitoyl-2-linoleoylphosphatidylethanolamine + H2O
-
 Rattus norvegicus N-octanoylethanolamine + 1-palmitoyl-2-linoleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-oleoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-oleoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Homo sapiens N-palmitoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-palmitoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-palmitoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?
3.1.4.54 N-stearoyl-1,2-dioleoyl-phosphatidylethanolamine + H2O
-
 Rattus norvegicus N-stearoylethanolamine + 1,2-dioleoyl-sn-glycerol 3-phosphate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.1.4.4 ? x * 46000, SDS-PAGE Rattus norvegicus
3.1.4.54 ? x * 46000, SDS-PAGE Rattus norvegicus

Synonyms

EC Number Synonyms Comment Organism
3.1.4.4 NAPE-PLD
-
 Rattus norvegicus
3.1.4.54 anandamide-generating phospholipase D
-
 Rattus norvegicus