Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.21.83 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
Search for more literature for 3.4.21.83

Cloned(Commentary)

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

Protein Variants

Protein Variants Comment Organism
E607Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E607Q/E609Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609A the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609F the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609L the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609Q/E610Q the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
E609R the mutant shows reduced kcat/Km compared to the wild type enzyme Trypanosoma brucei
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

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

Inhibitors

Inhibitors Comment Organism Structure
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]

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

Molecular Weight [Da]

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

Organic Solvent Stability

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

Organism

Organism UniProt Comment Textmining
Trypanosoma brucei
-
-
-

Purification (Commentary)

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

Specific Activity [micromol/min/mg]

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

Substrates and Products (Substrate)

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

Subunits

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

Synonyms

Synonyms Comment Organism
oligopeptidase B
-
 Trypanosoma brucei
OPB
-
 Trypanosoma brucei

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
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]

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

kcat/KM [mM/s]

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