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Literature summary for 3.4.22.69 extracted from

  • Kuo, C.J.; Shih, Y.P.; Kan, D.; Liang, P.H.
    Engineering a novel endopeptidase based on SARS 3CL(pro) (2009), Biotechniques, 47, 1029-1032.
    View publication on PubMed

Cloned(Commentary)

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

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 [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/ Products (Substrates)

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

Organism UniProt Comment Textmining
Severe acute respiratory syndrome-related coronavirus
-
-
-

Purification (Commentary)

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 and Products (Substrate)

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

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 [1/s]

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

General Information Comment Organism
physiological function 3CLpro is required for viral replication Severe acute respiratory syndrome-related coronavirus