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Literature summary for 3.4.21.83 extracted from

  • Ovchinnikova, M.V.; Mikhailova, A.G.; Karlinsky, D.M.; Gorlenko, V.A.; Rumsh, L.D.
    Reversible cyclic thermal inactivation of oligopeptidase B from Serratia proteamaculans (2018), Acta Naturae, 10, 65-70 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.21.83

Cloned(Commentary)

Cloned (Comment) Organism
recombinant expression of wild-type and mutant enzymes in Escherichia coli strain BL21(DE3) Serratia proteamaculans

Protein Variants

Protein Variants Comment Organism
D647A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 2fold lower Serratia proteamaculans
D649A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
E125A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 20-25% higher than that of the wild-type enzyme Serratia proteamaculans
E75A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 20-25% higher than that of the wild-type enzyme Serratia proteamaculans
H652X site-directed mutagenesis Serratia proteamaculans
K655A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
K660A site-directed mutagenesis, without Ca2+, the thermal stability of the mutant is 1.5-2fold lower Serratia proteamaculans
additional information calcixadum ions are a destabilizing factor for all PSP mutant variants. At 50 mM Ca2+, the difference in the thermal stability of all PSP variants is less marked, but the E75A mutant is the most thermally labile one, detailed overview. Substitution of the corresponding charged amino acid residues for the unxadcharged ones in OpdB from Trypanosoma brucei significantly rexadduces the thermal stability of these mutants Serratia proteamaculans

Metals/Ions

Metals/Ions Comment Organism Structure
Ca2+ calcium ions impede the reverse transition to the closed form. The reduction in the thermal stability of PSP in the presence of Ca2+ can be attributed to the destruction of the salt bridges SB2 and SB3 that takes place as Ca2+ binds to the E494 and D460 residues parxadtaking in bridge formation. Calcixadum ions are a destabilizing factor for all PSP mutant variants, overview Serratia proteamaculans

Organism

Organism UniProt Comment Textmining
Serratia proteamaculans A8GHH5
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-
Serratia proteamaculans 568 A8GHH5
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-

Purification (Commentary)

Purification (Comment) Organism
recombinant wild-type and mutant enzymes from Escherichia coli strain Bl21(DE3) Serratia proteamaculans

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
Nalpha-benzoyl-DL-argininyl-4-nitroanilide + H2O i.e. BAPNA, substrate of wild-type enzyme PSP, lower activity with truncated enzyme PSP-Chtr Serratia proteamaculans Nalpha-benzoyl-DL-arginine + 4-nitroaniline
-
 ?
Nalpha-benzoyl-DL-argininyl-4-nitroanilide + H2O i.e. BAPNA, substrate of wild-type enzyme PSP, lower activity with truncated enzyme PSP-Chtr Serratia proteamaculans 568 Nalpha-benzoyl-DL-arginine + 4-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
OpdB
-
 Serratia proteamaculans
PSP
-
 Serratia proteamaculans
Spro_3467
-
 Serratia proteamaculans

Temperature Optimum [°C]

Temperature Optimum [°C] Temperature Optimum Maximum [°C] Comment Organism
25
-
 assay at Serratia proteamaculans

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
37
-
 wild-type and mutant PSP enzymes can undergo reversible thermal inactivation at 37°C, pH 8.0, with activity being restored or even increased with respect to the initial one upon subsequent cooling. The process can be repeated several times, with the same results achieved (up to 5 cycles). This effect can be explained by a shift in the equilibrium between the inactive open form of the enzyme and the active closed one upon variation of the incubation temperature, overview Serratia proteamaculans

pH Optimum

pH Optimum Minimum pH Optimum Maximum Comment Organism
8
-
 assay at Serratia proteamaculans

General Information

General Information Comment Organism
evolution oligopeptidase B (OpdB) is a trypsin-like serine peptidase belonging to the prolyl oligopeptidase family Serratia proteamaculans
malfunction substitution of the corresponding charged amino acid residues for the unxadcharged ones in OpdB from Trypanosoma brucei significantly rexadduces the thermal stability of these mutants Serratia proteamaculans