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Literature summary extracted from

  • Guerin, M.E.; Kordulakova, J.; Schaeffer, F.; Svetlikova, Z.; Buschiazzo, A.; Giganti, D.; Gicquel, B.; Mikusova, K.; Jackson, M.; Alzari, P.M.
    Molecular recognition and interfacial catalysis by the essential phosphatidylinositol mannosyltransferase PimA from mycobacteria (2007), J. Biol. Chem., 282, 20705-20714.
    View publication on PubMed

Activating Compound

EC Number Activating Compound Comment Organism Structure
2.4.1.345 additional information PimA attaches to the membrane through its N-terminal domain and this association leads to enzyme activation Mycolicibacterium smegmatis
2.4.1.345 phospholipid aggregates the protein is able to interact with mono-disperse phosphatidylinositol through its active site cleft and also with phospholipid aggregates (micelles or liposomes), possibly through a different region of the protein. The latter interactions stimulate the catalytic activity Mycolicibacterium smegmatis

Cloned(Commentary)

EC Number Cloned (Comment) Organism
2.4.1.345
-
 Mycolicibacterium smegmatis
2.4.1.345 expressed in Escherichia coli Mycolicibacterium smegmatis

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
2.4.1.345 crystal structures of PimA in complex with GDP and GDP-Man are determined using multiplewavelength anomalous diffraction methods at 2.4 A and 2.6 A resolution respectively Mycolicibacterium smegmatis
2.4.1.345 crystal structures of PimA in complex with GDP and GDP-Man is determined using multiple-wavelength anomalous diffraction methods at 2.4 and 2.6 A of resolution, respectively Mycolicibacterium smegmatis
2.4.1.345 in complex with GDP and GDP-Man, to 2.4 and 2.6 A resolution, respectively. The structure of PimA in complex with GDP-mannose shows the two-domain organization and the catalytic machinery typical of GT-B glycosyltransferases. Model wherein PimA attaches to the membrane through its N-terminal domain, and this association leads to enzyme activation Mycolicibacterium smegmatis

Protein Variants

EC Number Protein Variants Comment Organism
2.4.1.345 DELTA59-70 mutation of the beta3-beta2 loop: mutant enzyme is still able to bind GDP with affinities in the submicromolar but PimA is completely inactivated and the ability of the protein to bind phospholipid aggregates is drastically impaired Mycolicibacterium smegmatis
2.4.1.345 E274A mutation results in complete enzyme inactivation Mycolicibacterium smegmatis
2.4.1.345 H118A mutation results in complete enzyme inactivation Mycolicibacterium smegmatis
2.4.1.345 additional information a PimA mutant in which the beta3-alpha2 loop is deleted by mutagenesis (PimA59–70) is still able to bind GDP with affinities in the submicromolar range but inactive and impaired the abilityto bind phospholipid aggregates Mycolicibacterium smegmatis
2.4.1.345 R201A mutation results in complete enzyme inactivation Mycolicibacterium smegmatis
2.4.1.345 R77S/K78S/K80S/K81S mutation of the four basic residues on alpha-helix 2: mutant enzyme is still able to bind GDP with affinities in the submicromolar but PimA is completely inactivated and the ability of the protein to bind phospholipid aggregates is drastically impaired Mycolicibacterium smegmatis
2.4.1.345 R77S/K78S/K80S/K81S mutant is still able to bind GDP with affinities in the submicromolar range but inactive and impaired the ability to bind phospholipid aggregates Mycolicibacterium smegmatis
2.4.1.345 T126W ability to produce phosphatidylinositol monomannoside (PIM1) is retained, enzymatic activity is similar to wild-type Mycolicibacterium smegmatis
2.4.1.345 T9A mutation results in complete enzyme inactivation Mycolicibacterium smegmatis

Localization

EC Number Localization Comment Organism GeneOntology No. Textmining
2.4.1.345 membrane PimA attaches to the membrane through its N-terminal domain and this association leads to enzyme activation Mycolicibacterium smegmatis 16020
-

Natural Substrates/ Products (Substrates)

EC Number Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
2.4.1.345 GDP-mannose + phosphatidylinositol Mycolicibacterium smegmatis PimA is responsible for the initial mannosylation of phosphatidylinositol GDP + phosphatidyl-(2-O-alpha-D-manno-pyranosyl)-myo-inositol
-
 ?

Organism

EC Number Organism UniProt Comment Textmining
2.4.1.345 Mycolicibacterium smegmatis
-
-
-
2.4.1.345 Mycolicibacterium smegmatis A0QWG6
-
-
2.4.1.345 Mycolicibacterium smegmatis ATCC 700084 A0QWG6
-
-

Reaction

EC Number Reaction Comment Organism Reaction ID
2.4.1.345 GDP-alpha-D-mannopyranose + 1-acyl-3-(2-(6-O-acyl-alpha-D-mannopyranosyl)-1D-myo-inositolphospho)-2-acyl-sn-glycerol = GDP + 1-acyl-3-(2-(6-O-acyl-alpha-D-mannopyranosyl)-6-O-alpha-D-mannopyranosyl-1D-myo-inositolphospho)-2-acyl-sn-glycerol based on structural, calorimetric, and mutagenesis studies, a model is proposed wherein PimA attaches to the membrane through its N-terminal domain, and this association leads to enzyme activation Mycolicibacterium smegmatis
a

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
2.4.1.345 additional information
-
 enzymatic activity is significantly increased using 1,2-dioctanoyl-sn-glycero-3-phospho-inositol as a substrate Mycolicibacterium smegmatis

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2.4.1.345 GDP-mannose + 1,2-dioctanoyl-sn-glycero-3-phosphoinositol
-
 Mycolicibacterium smegmatis ?
-
 ?
2.4.1.345 GDP-mannose + phosphatidylinositol
-
 Mycolicibacterium smegmatis GDP + phosphatidyl-(2-O-alpha-D-manno-pyranosyl)-myo-inositol
-
 ?
2.4.1.345 GDP-mannose + phosphatidylinositol PimA is responsible for the initial mannosylation of phosphatidylinositol Mycolicibacterium smegmatis GDP + phosphatidyl-(2-O-alpha-D-manno-pyranosyl)-myo-inositol
-
 ?
2.4.1.345 GDP-mannose + phosphatidylinositol
-
 Mycolicibacterium smegmatis GDP + phosphatidyl-(2-O-alpha-D-mannopyranosyl)-myo-inositol
-
 ?

Synonyms

EC Number Synonyms Comment Organism
2.4.1.345 PimA
-
 Mycolicibacterium smegmatis

Temperature Optimum [°C]

EC Number Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
2.4.1.345 37
-
 assay at Mycolicibacterium smegmatis

pH Optimum

EC Number pH Optimum Minimum pH Optimum Maximum Comment Organism
2.4.1.345 7.5
-
 assay at Mycolicibacterium smegmatis

General Information

EC Number General Information Comment Organism
2.4.1.345 physiological function the stoichiometry of the enzyme-substrate complex strongly depends on phosphatidylinositol concentration. The protein is able to interact with mono-disperse phosphatidylinositol through its active site cleft and also with phospholipid aggregates (micelles or liposomes), possibly through a different region of the protein. The latter interactions stimulate the catalytic activity Mycolicibacterium smegmatis