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Literature summary extracted from

  • Ashraf, M.; Sreejith, P.; Yadav, U.; Komath, S.S.
    Catalysis by N-acetyl-D-glucosaminylphosphatidylinositol de-N-acetylase (PIG-L) from Entamoeba histolytica: new roles for conserved residues (2013), J. Biol. Chem., 288, 7590-7595.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.5.1.89 recombinant expression of wild-type and mutant N-terminally MBP-tagged cytoplasmic catalytic domains in Escherichia coli strain TB1 Entamoeba histolytica

Protein Variants

EC Number Protein Variants Comment Organism
3.5.1.89 D102A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 D133A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 D45A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 D46A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is only slightly active in absence of metal ions and not stimulated by metal ions Entamoeba histolytica
3.5.1.89 D47A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is only slightly active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 E79A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 H140A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is only slightly active in absence of metal ions and not stimulated by metal ions Entamoeba histolytica
3.5.1.89 H143A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is only slightly active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 H43A site-directed mutagenesis of the isolated cytoplasmic catalytic domain, the mutant is only slightly active in absence of metal ions, but well stimulated by metal ions Entamoeba histolytica
3.5.1.89 additional information comparison of secondary conformation of the mutant proteins, overview Entamoeba histolytica

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.5.1.89 0.00137
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant H140A cytoplasmic catalytic domain, metal added Entamoeba histolytica
3.5.1.89 0.0016
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant H140A cytoplasmic catalytic domain, no metal added Entamoeba histolytica
3.5.1.89 0.00182
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant D46A cytoplasmic catalytic domain, no metal added Entamoeba histolytica
3.5.1.89 0.00195
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant D46A cytoplasmic catalytic domain, metal added Entamoeba histolytica
3.5.1.89 0.00195
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant wild-type cytoplasmic catalytic domain, no metal added Entamoeba histolytica
3.5.1.89 0.00199
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant H43A cytoplasmic catalytic domain, no metal added Entamoeba histolytica
3.5.1.89 0.00209
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant wild-type cytoplasmic catalytic domain, metal added Entamoeba histolytica
3.5.1.89 0.00263
-
 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol pH 5.5, 37°C, recombinant mutant H43A cytoplasmic catalytic domain, metal added Entamoeba histolytica

Metals/Ions

EC Number Metals/Ions Comment Organism Structure
3.5.1.89 Mg2+ stimulates Entamoeba histolytica
3.5.1.89 Mn2+ stimulates, binding of Mn2+ to the isolated cytoplasmic catalytic domain of the enzyme results in a significant alteration in the global conformation of the protein Entamoeba histolytica
3.5.1.89 additional information the enzyme exhibits a novel metal-independent albeit metal-stimulated activity Entamoeba histolytica

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
3.5.1.89 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + H2O Entamoeba histolytica
-
 6-(alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + acetate
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
3.5.1.89 Entamoeba histolytica C4M0W5
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.5.1.89 recombinant N-terminally wild-type and mutant MBP-tagged cytoplasmic catalytic domains from Escherichia coli strain TB1 by amylose affinity chromatography and removal of the tag, although the MBP tag does not significantly alter the activity of the enzyme Entamoeba histolytica

Reaction

EC Number Reaction Comment Organism Reaction ID
3.5.1.89 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + H2O = 6-(alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + acetate a general acid-base pair catalytic mechanism that involves enzyme residues Asp46 and His140 Entamoeba histolytica
a

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.5.1.89 6-(N-acetyl-alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + H2O
-
 Entamoeba histolytica 6-(alpha-D-glucosaminyl)-1-phosphatidyl-1D-myo-inositol + acetate
-
 ?

Synonyms

EC Number Synonyms Comment Organism
3.5.1.89 N-acetyl-D-glucosaminylphosphatidylinositol de-N-acetylase
-
 Entamoeba histolytica
3.5.1.89 PIG-L
-
 Entamoeba histolytica

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
3.5.1.89 37
-
 assay at Entamoeba histolytica

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
3.5.1.89 5.5
-
 assay at Entamoeba histolytica

General Information

EC Number General Information Comment Organism
3.5.1.89 metabolism the enzyme functions at the second step of GPI anchor biosynthesis, converting N-acetylglucosaminylphosphatidylinositol to glucosaminylphosphatidylinositol. This step is conserved in the GPI biosynthesis pathway Entamoeba histolytica
3.5.1.89 additional information residues Asp46 and His140 of the enzyme are important for catalysis Entamoeba histolytica