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Literature summary for 3.4.22.69 extracted from

  • Barrila, J.; Gabelli, S.B.; Bacha, U.; Amzel, L.M.; Freire, E.
    Mutation of Asn28 disrupts the dimerization and enzymatic activity of SARS 3CLpro (2010), Biochemistry, 49, 4308-4317.
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.69

Cloned(Commentary)

Cloned (Comment) Organism
expressed in Escherichia coli BL21 Star(DE3) cells Severe acute respiratory syndrome-related coronavirus

Crystallization (Commentary)

Crystallization (Comment) Organism
mutant enzyme N28A, hanging drop vapor diffusion method, using 1.0 M sodium malonate (pH 7.0), and 5-6% (v/v) isopropanol Severe acute respiratory syndrome-related coronavirus

Protein Variants

Protein Variants Comment Organism
N28A the mutation almost completely inactivates the enzyme and results in decreased dimerization Severe acute respiratory syndrome-related coronavirus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.05642
-
 Dabcyl-KTSAVLQSGFRKME-EDANS wild type enzyme, in 10 mM sodium phosphate, 10 mM sodium chloride, 1 mM EDTA, 1 mM TCEP, pH 7.4, at 25°C Severe acute respiratory syndrome-related coronavirus
0.06783
-
 Dabcyl-KTSAVLQSGFRKME-EDANS mutant enzyme N28A, in 10 mM sodium phosphate, 10 mM sodium chloride, 1 mM EDTA, 1 mM TCEP, pH 7.4, at 25°C Severe acute respiratory syndrome-related coronavirus

Molecular Weight [Da]

Molecular Weight [Da] Molecular Weight Maximum [Da] Comment Organism
34000
-
 2 * 34000 Severe acute respiratory syndrome-related coronavirus

Natural Substrates/ Products (Substrates)

Natural Substrates Organism Comment (Nat. Sub.) Natural Products Comment (Nat. Pro.) Rev. Reac.
Dabcyl-KTSAVLQSGFRKME-EDANS + H2O Severe acute respiratory syndrome-related coronavirus
-
 Dabcyl-KTSAVLQ + SGFRKME-EDANS
-
 ?
Dabcyl-KTSAVLQSGFRKME-EDANS + H2O Severe acute respiratory syndrome-related coronavirus Tor2
-
 Dabcyl-KTSAVLQ + SGFRKME-EDANS
-
 ?

Organism

Organism UniProt Comment Textmining
Severe acute respiratory syndrome-related coronavirus
-
-
-
Severe acute respiratory syndrome-related coronavirus Tor2
-
-
-

Purification (Commentary)

Purification (Comment) Organism
nickel affinity column chromatography and Q-Sepharose column chromatography, and Superdex 75 gel filtration 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-KTSAVLQ + SGFRKME-EDANS
-
 ?
Dabcyl-KTSAVLQSGFRKME-EDANS + H2O
-
 Severe acute respiratory syndrome-related coronavirus Tor2 Dabcyl-KTSAVLQ + SGFRKME-EDANS
-
 ?

Subunits

Subunits Comment Organism
homodimer 2 * 34000 Severe acute respiratory syndrome-related coronavirus

Synonyms

Synonyms Comment Organism
coronavirus main protease
-
 Severe acute respiratory syndrome-related coronavirus
SARS 3CLpro
-
 Severe acute respiratory syndrome-related coronavirus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.012
-
 Dabcyl-KTSAVLQSGFRKME-EDANS mutant enzyme N28A, in 10 mM sodium phosphate, 10 mM sodium chloride, 1 mM EDTA, 1 mM TCEP, pH 7.4, at 25°C Severe acute respiratory syndrome-related coronavirus
0.643
-
 Dabcyl-KTSAVLQSGFRKME-EDANS wild type enzyme, in 10 mM sodium phosphate, 10 mM sodium chloride, 1 mM EDTA, 1 mM TCEP, pH 7.4, at 25°C Severe acute respiratory syndrome-related coronavirus