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Literature summary extracted from

  • McMullen, T.W.; Li, J.; Sheffield, P.J.; Aoki, J.; Martin, T.W.; Arai, H.; Inoue, K.; Derewenda, Z.S.
    The functional implications of the dimerization of the catalytic subunits of the mammalian brain platelet-activating factor acetylhydrolase (Ib) (2000), Protein Eng., 13, 865-871.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.1.1.47 expression of the catalytic subunit alpha1-homodimer and alpha1-alpha2-heterodimer in Escherichia coli as GST-fusion protein, expression of wild-type and mutant enzymes as GST-fusion proteins in Escherichia coli Bos taurus

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
3.1.1.47 purified recombinant R22K mutant, X-ray structure determination at 1.2 A resolution Bos taurus

Protein Variants

EC Number Protein Variants Comment Organism
3.1.1.47 D20N site-directed mutagenesis, mutant is not efficiently expressed Bos taurus
3.1.1.47 D20R site-directed mutagenesis, mutant is not efficiently expressed Bos taurus
3.1.1.47 L194A site-directed mutagenesis, decreased Km and Vmax compared to the wild-type enzyme Bos taurus
3.1.1.47 L26A site-directed mutagenesis, 60% reduced activity compared to the wild-type enzyme Bos taurus
3.1.1.47 L48A site-directed mutagenesis, decreased Km and Vmax compared to the wild-type enzyme Bos taurus
3.1.1.47 R22E site-directed mutagenesis, mutant is not efficiently expressed Bos taurus
3.1.1.47 R22K site-directed mutagenesis, inactive mutant Bos taurus
3.1.1.47 R29K site-directed mutagenesis, slightly reduced activity compared to the wild-type enzyme Bos taurus
3.1.1.47 T103S site-directed mutagenesis, decreased Km and Vmax compared to the wild-type enzyme Bos taurus

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.1.1.47 additional information
-
additional information kinetics Bos taurus
3.1.1.47 0.082
-
acetylated platelet-activation factor mutant T103S, pH 7.4, 37°C Bos taurus
3.1.1.47 0.092
-
acetylated platelet-activation factor mutant L48A, pH 7.4, 37°C Bos taurus
3.1.1.47 0.135
-
acetylated platelet-activation factor mutant L194A, pH 7.4, 37°C Bos taurus
3.1.1.47 0.206
-
acetylated platelet-activation factor wild-type enzyme, pH 7.4, 37°C Bos taurus

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.1.1.47 Ca2+ influences the dimerization of the enzyme Bos taurus

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.1.1.47 26000
-
isozyme Ib, 2 * 26000, subunits alpha1 and/or subunit alpha2, + 1 * 45000, beta-subunit Bos taurus
3.1.1.47 45000
-
isozyme Ib, 2 * 26000, subunits alpha1 and/or subunit alpha2, + 1 * 45000, beta-subunit Bos taurus

Organism

EC Number Organism UniProt Comment Textmining
3.1.1.47 Bos taurus
-
isoform Ib
-

Reaction

EC Number Reaction Comment Organism Reaction ID
3.1.1.47 1-alkyl-2-acetyl-sn-glycero-3-phosphocholine + H2O = 1-alkyl-sn-glycero-3-phosphocholine + acetate Arg29 and Arg22 in the one monomer contribute to the catalytic competence of the active site across the dimer interface, and complement the catalytic triad of Ser47, Asp192, and His195 in the second monomer Bos taurus

Source Tissue

EC Number Source Tissue Comment Organism Textmining
3.1.1.47 brain isoform Ib Bos taurus
-

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.1.1.47 2-acetyl-1-alkyl-sn-glycero-3-phosphocholine + H2O i.e. platelet-activation factor Bos taurus 1-alkyl-sn-glycero-3-phosphocholine + acetate
-
?
3.1.1.47 acetylated platelet-activation factor + H2O
-
Bos taurus ?
-
?

Subunits

EC Number Subunits Comment Organism
3.1.1.47 More determination of structure-function relationship, dimerization of the brain isozyme is essential for both catalytic activity and stability, the monomeric enzyme is unstable, the alpha1 and alpha2 are catalytic subunits, the beta-subunit is a regulatory one Bos taurus
3.1.1.47 trimer isozyme Ib, 2 * 26000, subunits alpha1 and/or subunit alpha2, + 1 * 45000, beta-subunit Bos taurus

Synonyms

EC Number Synonyms Comment Organism
3.1.1.47 PAF-AH
-
Bos taurus
3.1.1.47 platelet-activating factor acetylhydrolase
-
Bos taurus

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.1.1.47 37
-
assay at Bos taurus

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.1.1.47 47.5
-
unfolding and irreversible denaturation at pH 9.5 Bos taurus
3.1.1.47 52.3
-
unfolding and irreversible denaturation at pH 6.5 Bos taurus

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.1.1.47 7.4
-
assay at Bos taurus

pH Range

EC Number pH Minimum pH Maximum Comment Organism
3.1.1.47 additional information
-
-
Bos taurus