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Literature summary extracted from

  • Mohd Ismail, N.I.; Yuasa, T.; Yuasa, K.; Nambu, Y.; Nisimoto, M.; Goto, M.; Matsuki, H.; Inoue, M.; Nagahama, M.; Tsuji, A.
    A critical role for highly conserved Glu(610) residue of oligopeptidase B from Trypanosoma brucei in thermal stability (2010), J. Biochem., 147, 201-211.
    View publication on PubMed

Cloned(Commentary)

EC Number Cloned (Comment) Organism
3.4.21.83 expressed in Escherichia coli BL21 cells Trypanosoma brucei

Protein Variants

EC Number Protein Variants Comment Organism
3.4.21.83 E607Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E607Q/E609Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609A the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609F the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609L the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609Q/E610Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E609R the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
3.4.21.83 E610Q the mutant shows reduced kcat/Km compared to the wild type enzyme, the mutation results in a marked decrease in thermal stability, the mutant is inactivated to 63% of the control by 3 M urea Trypanosoma brucei

General Stability

EC Number General Stability Organism
3.4.21.83 the wild type enzyme is not digested by trypsin at 25°C Trypanosoma brucei

Inhibitors

EC Number Inhibitors Comment Organism Structure
3.4.21.83 additional information reducing reagent such as dithiothreitol has no effect on the inactivation of wild type and mutant OPBs by heat and urea treatment Trypanosoma brucei

KM Value [mM]

EC Number KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
3.4.21.83 0.00268
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei
3.4.21.83 0.00334
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei
3.4.21.83 0.00525
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 0.00528
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei
3.4.21.83 0.00568
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 0.00586
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 0.00637
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 0.0069
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 0.00728
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 0.00807
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei
3.4.21.83 0.00904
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 0.00913
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 0.0101
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 0.0113
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 0.0127
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 0.0127
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 0.015
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 0.0173
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 0.0184
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei
3.4.21.83 0.0215
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei

Molecular Weight [Da]

EC Number Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
3.4.21.83 84000
-
 2 * 84000, calculated from amino acid sequence Trypanosoma brucei
3.4.21.83 160000
-
 gel filtration Trypanosoma brucei

Organic Solvent Stability

EC Number Organic Solvent Comment Organism
3.4.21.83 urea the wild type enzyme is resistant to treatment with 3 M urea Trypanosoma brucei

Organism

EC Number Organism UniProt Comment Textmining
3.4.21.83 Trypanosoma brucei
-
-
-

Purification (Commentary)

EC Number Purification (Comment) Organism
3.4.21.83 Ni-NTA agarose column chromatography Trypanosoma brucei

Specific Activity [micromol/min/mg]

EC Number Specific Activity Minimum [µmol/min/mg] Specific Activity Maximum [µmol/min/mg] Comment Organism
3.4.21.83 0.0001
-
 mutant enzyme E607Q/E609Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei
3.4.21.83 0.000323
-
 mutant enzyme E607Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei
3.4.21.83 0.011
-
 mutant enzyme E609Q/E610Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei
3.4.21.83 0.0209
-
 mutant enzyme E610Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei
3.4.21.83 0.0221
-
 mutant enzyme E609Q, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei
3.4.21.83 0.0279
-
 wild type enzyme, towards benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin, at 37°C Trypanosoma brucei

Substrates and Products (Substrate)

EC Number Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
3.4.21.83 adrenocorticotropic hormone + H2O
-
 Trypanosoma brucei SYSMEHFR + WGK + WGKPVGK + VYP
-
 ?
3.4.21.83 benzoyl-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Trypanosoma brucei benzoyl-L-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.21.83 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Trypanosoma brucei benzyloxycarbonyl-L-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.21.83 benzyloxycarbonyl-L-Phe-L-Arg-7-amido-4-methylcoumarin + H2O
-
 Trypanosoma brucei benzyloxycarbonyl-L-Phe-L-Arg + 7-amino-4-methylcoumarin
-
 ?
3.4.21.83 Glucagon + H2O wild type enzyme cleaves Lys12-Tyr13, Arg17-Arg18, and Arg18-Ala19 bonds of glucagon Trypanosoma brucei ?
-
 ?

Subunits

EC Number Subunits Comment Organism
3.4.21.83 homodimer 2 * 84000, calculated from amino acid sequence Trypanosoma brucei

Synonyms

EC Number Synonyms Comment Organism
3.4.21.83 oligopeptidase B
-
 Trypanosoma brucei
3.4.21.83 OPB
-
 Trypanosoma brucei

Temperature Stability [°C]

EC Number Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
3.4.21.83 45 53 following treatment at 45°C for 20 min the relative activity of wild type enzyme is 89%, the melting temperature of the wild type enzyme is at 53.3°C Trypanosoma brucei

Turnover Number [1/s]

EC Number Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3.4.21.83 0.758
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 1.24
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 1.42
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei
3.4.21.83 1.46
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 1.58
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 1.64
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 1.77
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 4.98
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 5.03
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 5.37
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei
3.4.21.83 5.42
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 6.84
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 7.81
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei
3.4.21.83 8.29
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei
3.4.21.83 9.36
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 9.53
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 11.2
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 13.3
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei
3.4.21.83 15.2
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 15.8
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei

kcat/KM [mM/s]

EC Number kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
3.4.21.83 50
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 67
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 77
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei
3.4.21.83 83
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q, at 37°C Trypanosoma brucei
3.4.21.83 129
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 217
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E607Q/E609Q, at 37°C Trypanosoma brucei
3.4.21.83 250
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609R, at 37°C Trypanosoma brucei
3.4.21.83 265
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609F, at 37°C Trypanosoma brucei
3.4.21.83 551
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 647
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 677
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 750
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q/E610Q, at 37°C Trypanosoma brucei
3.4.21.83 790
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609L, at 37°C Trypanosoma brucei
3.4.21.83 1030
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609A, at 37°C Trypanosoma brucei
3.4.21.83 1480
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei
3.4.21.83 1650
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 1960
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E609Q, at 37°C Trypanosoma brucei
3.4.21.83 2480
-
 benzoyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei
3.4.21.83 2590
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin mutant enzyme E610Q, at 37°C Trypanosoma brucei
3.4.21.83 4960
-
 benzyloxycarbonyl-L-Arg-7-amido-4-methylcoumarin wild type enzyme, at 37°C Trypanosoma brucei