Cloned (Comment) | Organism |
---|---|
expressed in Escherichia BL21 Star (DE3) cells | Clostridioides difficile |
Crystallization (Comment) | Organism |
---|---|
- |
Clostridioides difficile |
Protein Variants | Comment | Organism |
---|---|---|
E143A/Y178F | inactive | Clostridioides difficile |
E184A | the mutant shows reduced activity for Abz-DVVNPPVPPSKDD-CONH2 (15%) and Abz-DVVNAPVPPSKDD-CONH2 (9%) and residual activity for Abz-DVVNPAVPPSKDD-CONH2 (1.2%) compared to the wild type enzyme | Clostridioides difficile |
E184K | the mutant shows residual activity compared to the wild type enzyme | Clostridioides difficile |
K101A | the mutant shows dramatically reduced activity compared to the wild type enzyme | Clostridioides difficile |
K101E | the mutant shows dramatically reduced activity compared to the wild type enzyme | Clostridioides difficile |
K101R | the mutant shows about wild type activity | Clostridioides difficile |
W103A | inactive | Clostridioides difficile |
W103F | the mutant shows residual activity for Abz-DVVNPPVPPSKDD-CONH2 compared to the wild type enzyme | Clostridioides difficile |
W103H | the mutant shows residual activity for Abz-DVVNPPVPPSKDD-CONH2 compared to the wild type enzyme | Clostridioides difficile |
W103Y | the mutant shows severely reduced activity for Abz-DVVNPPVPPSKDD-CONH2 and residual activity for Abz-DVVNPAVPPSKDD-CONH2 and Abz-DVVNAPVPPSKDD-CONH2 compared to the wild type enzyme | Clostridioides difficile |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0436 | - |
Abz-DVVNAPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
0.0825 | - |
Abz-DVVNPAVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
0.1366 | - |
Abz-DVVNPPVPPSKDD-CONH2 | mutant enzyme E184A, at pH 7.5 and 20°C | Clostridioides difficile | |
0.137 | - |
Abz-DVVNPPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile |
Metals/Ions | Comment | Organism | Structure |
---|---|---|---|
Zn2+ | required | Clostridioides difficile |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
Ac-EPNAAVP-CONH2 + H2O | Clostridioides difficile | heat shock protein 90-derived peptide | Ac-EPNA + AVP-CONH2 | - |
? | |
Ac-EPNAAVP-CONH2 + H2O | Clostridioides difficile 630 | heat shock protein 90-derived peptide | Ac-EPNA + AVP-CONH2 | - |
? | |
Ac-HLLPPPS-CONH2 + H2O | Clostridioides difficile | immunoglobulin A1-derived peptide | Ac-HLLP + PPS-CONH2 | - |
? | |
Ac-HLLPPPS-CONH2 + H2O | Clostridioides difficile 630 | immunoglobulin A1-derived peptide | Ac-HLLP + PPS-CONH2 | - |
? | |
Ac-SLRPAPP-CONH2 + H2O | Clostridioides difficile | fibrinogen beta-derived peptide | Ac-SLRP + APP-CONH2 | - |
? | |
adhesion molecule CD2831 + H2O | Clostridioides difficile | - |
? | - |
? | |
adhesion molecule CD2831 + H2O | Clostridioides difficile 630 | - |
? | - |
? | |
adhesion molecule CD3246 + H2O | Clostridioides difficile | - |
? | - |
? | |
adhesion molecule CD3246 + H2O | Clostridioides difficile 630 | - |
? | - |
? | |
additional information | Clostridioides difficile | the enzyme exhibits a unique specificity for Pro-Pro peptide bonds within the consensus sequence VNP-PVP. The enzyme prefers proline residues at P1'. The residues Lys-101, Trp-103, and Glu-184 are crucial for proteolytic activity | ? | - |
? | |
additional information | Clostridioides difficile 630 | the enzyme exhibits a unique specificity for Pro-Pro peptide bonds within the consensus sequence VNP-PVP. The enzyme prefers proline residues at P1'. The residues Lys-101, Trp-103, and Glu-184 are crucial for proteolytic activity | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Clostridioides difficile | Q183R7 | - |
- |
Clostridioides difficile 630 | Q183R7 | - |
- |
Purification (Comment) | Organism |
---|---|
immobilized metal-affinity chromatography and gel filtration | Clostridioides difficile |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
Abz-DVVNAPVPPSKDD-CONH2 + H2O | - |
Clostridioides difficile | Abz-DVVNA + PVPPSKDD-CONH2 | - |
? | |
Abz-DVVNPAVPPSKDD-CONH2 + H2O | - |
Clostridioides difficile | Abz-DVVNP + AVPPSKDD-CONH2 | - |
? | |
Abz-DVVNPPVPPSKDD-CONH2 + H2O | - |
Clostridioides difficile | Abz-DVVNP + PVPPSKDD-CONH2 | - |
? | |
Ac-EPNAAVP-CONH2 + H2O | heat shock protein 90-derived peptide | Clostridioides difficile | Ac-EPNA + AVP-CONH2 | - |
? | |
Ac-EPNAAVP-CONH2 + H2O | heat shock protein 90-derived peptide | Clostridioides difficile 630 | Ac-EPNA + AVP-CONH2 | - |
? | |
Ac-HLLPPPS-CONH2 + H2O | immunoglobulin A1-derived peptide | Clostridioides difficile | Ac-HLLP + PPS-CONH2 | - |
? | |
Ac-HLLPPPS-CONH2 + H2O | immunoglobulin A1-derived peptide | Clostridioides difficile 630 | Ac-HLLP + PPS-CONH2 | - |
? | |
Ac-SLRPAPP-CONH2 + H2O | fibrinogen beta-derived peptide | Clostridioides difficile | Ac-SLRP + APP-CONH2 | - |
? | |
adhesion molecule CD2831 + H2O | - |
Clostridioides difficile | ? | - |
? | |
adhesion molecule CD2831 + H2O | - |
Clostridioides difficile 630 | ? | - |
? | |
adhesion molecule CD3246 + H2O | - |
Clostridioides difficile | ? | - |
? | |
adhesion molecule CD3246 + H2O | - |
Clostridioides difficile 630 | ? | - |
? | |
additional information | the enzyme exhibits a unique specificity for Pro-Pro peptide bonds within the consensus sequence VNP-PVP. The enzyme prefers proline residues at P1'. The residues Lys-101, Trp-103, and Glu-184 are crucial for proteolytic activity | Clostridioides difficile | ? | - |
? | |
additional information | the enzyme exhibits a unique specificity for Pro-Pro peptide bonds within the consensus sequence VNP-PVP. The enzyme prefers proline residues at P1'. The residues Lys-101, Trp-103, and Glu-184 are crucial for proteolytic activity | Clostridioides difficile 630 | ? | - |
? |
Subunits | Comment | Organism |
---|---|---|
? | x * 24000, SDS-PAGE | Clostridioides difficile |
Synonyms | Comment | Organism |
---|---|---|
CD2830 | - |
Clostridioides difficile |
PPEP-1 | - |
Clostridioides difficile |
proline-proline endopeptidase-1 | - |
Clostridioides difficile |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.87 | - |
Abz-DVVNPAVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
1.17 | - |
Abz-DVVNPPVPPSKDD-CONH2 | mutant enzyme E184A, at pH 7.5 and 20°C | Clostridioides difficile | |
1.26 | - |
Abz-DVVNAPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
4.3 | - |
Abz-DVVNPPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile |
kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
8.565 | - |
Abz-DVVNPPVPPSKDD-CONH2 | mutant enzyme E184A, at pH 7.5 and 20°C | Clostridioides difficile | |
10.545 | - |
Abz-DVVNPAVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
28.899 | - |
Abz-DVVNAPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile | |
31.386 | - |
Abz-DVVNPPVPPSKDD-CONH2 | wild type enzyme, at pH 7.5 and 20°C | Clostridioides difficile |