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Literature summary extracted from

  • Jeyaharan, D.; Brackstone, C.; Schouten, J.; Davis, P.; Dixon, A.M.
    Characterisation of the carboxypeptidase G2 catalytic site and design of new inhibitors for cancer therapy (2018), ChemBioChem, 19, 1959-1968 .
    View publication on PubMed

Application

EC Number Application Comment Organism
3.4.17.11 pharmacology the enzyme is used in antibody directed enzyme prodrug therapy to catalyse the formation of an active drug from an inert prodrug. Free carboxypeptidase G2 in the bloodstream must be inhibited before administration of the prodrug in order to avoid a systemic reaction in the patient Pseudomonas sp. RS-16

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.17.11 expression in Escherichia coli BL21(DE3) Pseudomonas sp. RS-16

Protein Variants

EC Number Protein Variants Comment Organism
3.4.17.11 D141A substitution leads to a decrease in expression level (compared to wild type). Melting temperature is 53.2°C, compared to 60.4°C for wild-type enzyme Pseudomonas sp. RS-16
3.4.17.11 E175A substitution leads to a decrease in expression level (compared to wild type). Melting temperature is 71.4°C, compared to 60.4°C for wild-type enzyme. Mutation abolishes activity against methotrexate Pseudomonas sp. RS-16
3.4.17.11 E200A substitution leads to a decrease in expression level (compared to wild type). Melting temperature is 64.3°C, compared to 60.4°C for wild-type enzyme Pseudomonas sp. RS-16
3.4.17.11 H112A substitution leads to a high decrease in expression level (compared to wild type). Melting temperature is 28.2°C, compared to 60.4°C for wild-type enzyme Pseudomonas sp. RS-16
3.4.17.11 H385A substitution leads to a low decrease in expression level (compared to wild type). Melting temperature is 55.1°C, compared to 60.4°C for wild-type enzyme Pseudomonas sp. RS-16
3.4.17.11 K208A substitution leads to a high decrease in expression level (compared to wild type). Melting temperature is 53.4°C, compared to 60.4°C for wild-type enzyme. Mutation abolishes activity against methotrexate Pseudomonas sp. RS-16
3.4.17.11 S210A substitution leads to a high decrease in expression level (compared to wild type). Melting temperature is 54.4C, compared to 60.4°C for wild-type enzyme. Mutation abolishes activity against methotrexate Pseudomonas sp. RS-16
3.4.17.11 T357A substitution leads to a decrease in expression level (compared to wild type). Melting temperature is 55.4°C, compared to 60.4°C for wild-type enzyme. Mutation abolishes activity against methotrexate Pseudomonas sp. RS-16

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.17.11 (2R)-2-[(3-aminonaphthalene-2-carbonyl)amino]pentanedioic acid CP06 Pseudomonas sp. RS-16
3.4.17.11 (2R)-2-[[3-(2-methylpropanamido)naphthalene-2-carbonyl]amino]pentanedioic acid CP67 Pseudomonas sp. RS-16

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.17.11 52000
-
 SDS-PAGE Pseudomonas sp. RS-16

Organism

EC Number Organism UniProt Comment Textmining
3.4.17.11 Pseudomonas sp. RS-16 P06621
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.17.11
-
 Pseudomonas sp. RS-16

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.17.11 folic acid + H2O
-
 Pseudomonas sp. RS-16 pteroic acid + glutamic acid
-
 ?
3.4.17.11 methotrexate + H2O
-
 Pseudomonas sp. RS-16 4-[(2,4-diamino-6-(pteridinyl)methyl)-methylamino]-benzoic acid + L-glutamate
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.17.11 ? x * 52000, SDS-PAGE Pseudomonas sp. RS-16

Synonyms

EC Number Synonyms Comment Organism
3.4.17.11 carboxypeptidase G2
-
 Pseudomonas sp. RS-16
3.4.17.11 CPG2
-
 Pseudomonas sp. RS-16

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.17.11 28.2
-
 melting temperature of mutant enzyme H112A Pseudomonas sp. RS-16
3.4.17.11 53.2
-
 melting temperature of mutant enzyme D141A Pseudomonas sp. RS-16
3.4.17.11 53.4
-
 melting temperature of mutant enzyme K208A Pseudomonas sp. RS-16
3.4.17.11 54.4
-
 melting temperature of mutant enzyme S210A Pseudomonas sp. RS-16
3.4.17.11 55.1
-
 melting temperature of mutant enzyme H385A Pseudomonas sp. RS-16
3.4.17.11 55.4
-
 melting temperature of mutant enzyme T357A Pseudomonas sp. RS-16
3.4.17.11 60.4
-
 melting temperature of wild-type enzyme Pseudomonas sp. RS-16
3.4.17.11 64.3
-
 melting temperature of mutant enzyme E200A Pseudomonas sp. RS-16
3.4.17.11 71.4
-
 melting temperature of mutant enzyme E175A Pseudomonas sp. RS-16

IC50 Value

EC Number IC50 Value IC50 Value Maximum Comment Organism Inhibitor Structure
3.4.17.11 0.068
-
 pH 7.3, 37°C Pseudomonas sp. RS-16 (2R)-2-[(3-aminonaphthalene-2-carbonyl)amino]pentanedioic acid
3.4.17.11 0.113
-
 pH 7.3, 37°C Pseudomonas sp. RS-16 (2R)-2-[[3-(2-methylpropanamido)naphthalene-2-carbonyl]amino]pentanedioic acid