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Literature summary for 3.1.4.4 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)

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 activity comparable to wild-type Rattus norvegicus
C170S activity comparable to wild-type Rattus norvegicus
C222S activity comparable to wild-type Rattus norvegicus
C224S considerable reduction in activity Rattus norvegicus
C237S activity comparable to wild-type Rattus norvegicus
C255S activity comparable to wild-type Rattus norvegicus
C288S activity comparable to wild-type Rattus norvegicus
D147N complete loss of activity Rattus norvegicus
D189N less than 0.1% of wild-type activity Rattus norvegicus
D284N complete loss of activity Rattus norvegicus
D389N single nucleotide polymorphism, activity comparable to wild-type Homo sapiens
DELTAN138 complete loss of activity Rattus norvegicus
DELTAN55 activity comparable to wild-type Rattus norvegicus
DELTAN85 complete loss of activity Rattus norvegicus
H185N less than 0.1% of wild-type activity Rattus norvegicus
H187N less than 0.1% of wild-type activity Rattus norvegicus
H190N less than 0.1% of wild-type activity Rattus norvegicus
H253N complete loss of activity Rattus norvegicus
H321N complete loss of activity Rattus norvegicus
H331N 4% of wild-type activity Rattus norvegicus
H343N activity comparable to wild-type Rattus norvegicus
H353N activity comparable to wild-type Rattus norvegicus
H380R single nucleotide polymorphism, complete loss of activity Homo sapiens
H380R single nucleotide polymorphism, low catalytic activity Rattus norvegicus
L207F single nucleotide polymorphism, complete loss of activity Homo sapiens
L207F single nucleotide polymorphism, low catalytic activity Rattus norvegicus
S152A single nucleotide polymorphism, activity comparable to wild-type Homo sapiens

KM Value [mM]

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

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ 50% activation at 2.3 mM Rattus norvegicus
Mg2+ 50% activation at 2.3 mM Rattus norvegicus
Zn2+ catalytically active Zn2+ 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
-
N-acylethanolamine-specific phospholipase D, NAPE-PLD
-
Rattus norvegicus
-
N-acylethanolamine-specific phospholipase D, NAPE-PLD
-

Purification (Commentary)

Purification (Comment) Organism
recombinant enzyme Rattus norvegicus

Specific Activity [micromol/min/mg]

Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.016
-
37°C, pH 7.4, presence of 10 mM MgCl2, substrate N-palmitoyl-phosphatidylethanolamine Rattus norvegicus
2.4
-
37°C, pH 7.4, presence of 10 mM MgCl2, substrate N-lauroyl-phosphatidylethanolamine Rattus norvegicus

Storage Stability

Storage Stability Organism
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
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

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
glycerophospho-(N-palmitoyl)ethanolamine + H2O
-
Rattus norvegicus N-palmitoylethanolamine + ? 1% of the activity with N-palmitoyl-phosphatidylethanolamine ?
additional information no substrate: N-palmitoylethanolamine phosphate, phosphatidylcholine, phosphatidylserine, phosphatidylinositol Rattus norvegicus ?
-
?
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
-
?
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
-
?
N-lauroyl-phosphatidylethanolamine + H2O
-
Homo sapiens phosphatidic acid + N-lauroylethanolamine
-
?
N-lauroyl-phosphatidylethanolamine + H2O
-
Rattus norvegicus phosphatidic acid + N-lauroylethanolamine
-
?
N-palmitoyl-lyso-phosphatidylethanolamine + H2O
-
Rattus norvegicus N-palmitoylethanolamine + sn-glycerol 3-phosphate 4% of the activity with N-palmitoyl-phosphatidylethanolamine ?
N-palmitoyl-phosphatidylethanolamine + H2O
-
Homo sapiens phosphatidic acid + N-palmitoylethanolamine
-
?
N-palmitoyl-phosphatidylethanolamine + H2O
-
Rattus norvegicus phosphatidic acid + N-palmitoylethanolamine
-
?

Subunits

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

Synonyms

Synonyms Comment Organism
NAPE-PLD
-
Rattus norvegicus