Cloned (Comment) | Organism |
---|---|
expressed in Escherichia coli | Severe acute respiratory syndrome-related coronavirus |
wild type and His-tagged T25G mutant are expressed in Escherichia coli cells | Severe acute respiratory syndrome-related coronavirus |
Protein Variants | Comment | Organism |
---|---|---|
T25G | activity like wild-type (substrate DABCYL-Lys-Thr-Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys-Met-Glu-EDANS), higher specific activity than wild-type protein for substrate Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys | Severe acute respiratory syndrome-related coronavirus |
T25G | the mutant enzyme has an expanded S1' space that demonstrates 43.5-fold better kcat/Km compared with wild type in cleaving substrates with a larger Met at P1', mutant enzyme T25G shows a 12fold and 8fold higher activity against the substrates with Met and Leu at P1', respectively | Severe acute respiratory syndrome-related coronavirus |
T25S | almost complete loss of activity | Severe acute respiratory syndrome-related coronavirus |
KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.0186 | - |
SAVLQMGFRK | mutant enzyme T25G, at 37°C | Severe acute respiratory syndrome-related coronavirus | |
0.0186 | - |
Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys | T25G mutant protein | Severe acute respiratory syndrome-related coronavirus | |
0.0766 | - |
SAVLQMGFRK | wild type enzyme, at 37°C | Severe acute respiratory syndrome-related coronavirus | |
0.0766 | - |
Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys | wild-type protein | Severe acute respiratory syndrome-related coronavirus |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
additional information | Severe acute respiratory syndrome-related coronavirus | 3CLpro cleaves the replicase polyproteins at 11 sites with the conserved Gln-(Ser, Ala, Gly) sequences | ? | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Severe acute respiratory syndrome-related coronavirus | - |
- |
- |
Purification (Comment) | Organism |
---|---|
immobilized metal ion affinity chromatography (Ni2+) | Severe acute respiratory syndrome-related coronavirus |
Ni-NTA column chromatography | Severe acute respiratory syndrome-related coronavirus |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
dabcyl-KTSAVLQSGFRKME-EDANS + H2O | - |
Severe acute respiratory syndrome-related coronavirus | ? | - |
? | |
DABCYL-Lys-Thr-Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys-Met-Glu-EDANS + H2O | - |
Severe acute respiratory syndrome-related coronavirus | DABCYL-Lys-Thr-Ser-Ala-Val-Leu-Gln + Ser-Gly-Phe-Arg-Lys-Met-Glu-EDANS | - |
? | |
additional information | 3CLpro cleaves the replicase polyproteins at 11 sites with the conserved Gln-(Ser, Ala, Gly) sequences | Severe acute respiratory syndrome-related coronavirus | ? | - |
? | |
additional information | no cleavage of SAVLQPGFRK | Severe acute respiratory syndrome-related coronavirus | ? | - |
? | |
SAVLQEGFRK + H2O | the cleavage rate of the mutant enzyme T25G is remarkably higher compared to the wild type enzyme | Severe acute respiratory syndrome-related coronavirus | SAVLQ + EGFRK | - |
? | |
SAVLQFGFRK + H2O | the cleavage rate of the mutant enzyme T25G is remarkably higher compared to the wild type enzyme | Severe acute respiratory syndrome-related coronavirus | SAVLQ + FGFRK | - |
? | |
SAVLQGGFRK + H2O | the cleavage rate of the mutant enzyme T25G is similar to the wild type enzyme | Severe acute respiratory syndrome-related coronavirus | SAVLQ + GGFRK | - |
? | |
SAVLQHGFRK + H2O | low activity | Severe acute respiratory syndrome-related coronavirus | SAVLQ + HGFRK | - |
? | |
SAVLQKGFRK + H2O | low activity | Severe acute respiratory syndrome-related coronavirus | SAVLQ + KGFRK | - |
? | |
SAVLQLGFRK + H2O | the cleavage rate of the mutant enzyme T25G is remarkably higher compared to the wild type enzyme | Severe acute respiratory syndrome-related coronavirus | SAVLQ + LGFRK | - |
? | |
SAVLQMGFRK + H2O | the cleavage rate of the mutant enzyme T25G is remarkably higher compared to the wild type enzyme | Severe acute respiratory syndrome-related coronavirus | SAVLQ + MGFRK | - |
? | |
SAVLQSGFRK + H2O | best substrate for both, wild type and mutant enzyme T25G | Severe acute respiratory syndrome-related coronavirus | SAVLQ + SGFRK | - |
? | |
SAVLQWGFRK + H2O | low activity | Severe acute respiratory syndrome-related coronavirus | SAVLQ + WGFRK | - |
? | |
Ser-Ala-Val-Leu-Gln-Leu-Gly-Phe-Arg-Lys + H2O | substrate for T25G mutant protein | Severe acute respiratory syndrome-related coronavirus | Ser-Ala-Val-Leu-Gln + Leu-Gly-Phe-Arg-Lys | - |
? | |
Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys + H2O | - |
Severe acute respiratory syndrome-related coronavirus | Ser-Ala-Val-Leu-Gln + Met-Gly-Phe-Arg-Lys | - |
? | |
Ser-Ala-Val-Leu-Gln-Ser-Gly-Phe-Arg-Lys + H2O | - |
Severe acute respiratory syndrome-related coronavirus | Ser-Ala-Val-Leu-Gln + Ser-Gly-Phe-Arg-Lys | - |
? |
Synonyms | Comment | Organism |
---|---|---|
3C-like protease | - |
Severe acute respiratory syndrome-related coronavirus |
3cLpro | - |
Severe acute respiratory syndrome-related coronavirus |
SARS 3CLpro | - |
Severe acute respiratory syndrome-related coronavirus |
Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
---|---|---|---|---|---|
0.027 | - |
Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys | wild-type protein | Severe acute respiratory syndrome-related coronavirus | |
0.27 | - |
Ser-Ala-Val-Leu-Gln-Met-Gly-Phe-Arg-Lys | T25G mutant protein | Severe acute respiratory syndrome-related coronavirus | |
1.6 | - |
SAVLQMGFRK | wild type enzyme, at 37°C | Severe acute respiratory syndrome-related coronavirus | |
16.2 | - |
SAVLQMGFRK | mutant enzyme T25G, at 37°C | Severe acute respiratory syndrome-related coronavirus |
General Information | Comment | Organism |
---|---|---|
physiological function | 3CLpro is required for viral replication | Severe acute respiratory syndrome-related coronavirus |