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Literature summary extracted from

  • Sigala, P.; Caaveiro, J.; Ringe, D.; Petsko, G.; Herschlag, D.
    Hydrogen bond coupling in the ketosteroid isomerase active site (2009), Biochemistry, 48, 6932-6939.
    View publication on PubMedView publication on EuropePMC

Cloned(Commentary)

EC Number Cloned (Comment) Organism
5.3.3.1 expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha Comamonas testosteroni
5.3.3.1 expression of wild-type and mutant enzymes in Escherichia coli strain DH5alpha Pseudomonas putida

Crystallization (Commentary)

EC Number Crystallization (Comment) Organism
5.3.3.1 mutant D103N/D40N bound to inhibitor equilenin, hanging drop vapor diffusion method, 0.002 ml of 25 mg/ml protein with equilenin in a molar ratio of 1:1.2 in 40 mM potassium phosphate, pH 7.2, are mixed with 0.002 ml of reservoir solution conraining 1.4 M ammonium sulfate, and 6.5% v/v 2-propanol, pH 7.0, room temperature, 1 week, X-ray diffraction structure determination and analysis at 1.1 A resolution Pseudomonas putida

Protein Variants

EC Number Protein Variants Comment Organism
5.3.3.1 D103N/D40N site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the D103N mutation Pseudomonas putida
5.3.3.1 Y57F/D40N site-directed mutagenesis, 3,4,5-trifluorophenol bind as ionized phenolate to KSI containing the Y57F mutation Comamonas testosteroni

Inhibitors

EC Number Inhibitors Comment Organism Structure
5.3.3.1 equilenin reaction intermediate analogue, binds to the active site, binding structure, overview Comamonas testosteroni
5.3.3.1 equilenin reaction intermediate analogue, binds to the active site, binding structure, overview Pseudomonas putida

Organism

EC Number Organism UniProt Comment Textmining
5.3.3.1 Comamonas testosteroni
-
-
-
5.3.3.1 Pseudomonas putida P07445
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
5.3.3.1 recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity Comamonas testosteroni
5.3.3.1 recombinant wild-type and mutant enzymes from Escherichia coli strain DH5alpha to over 99% purity Pseudomonas putida

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
5.3.3.1 additional information mechanism of KSI-catalyzed steroid isomerization, direct detection of physical coupling between the Y16 and D103 hydrogen bonds, ligand binding, overview Pseudomonas putida ?
-
 ?
5.3.3.1 additional information mechanism of KSI-catalyzed steroid isomerization, overview Comamonas testosteroni ?
-
 ?

Synonyms

EC Number Synonyms Comment Organism
5.3.3.1 ketosteroid isomerase
-
 Comamonas testosteroni
5.3.3.1 ketosteroid isomerase
-
 Pseudomonas putida
5.3.3.1 KSI
-
 Comamonas testosteroni
5.3.3.1 KSI
-
 Pseudomonas putida