Any feedback?
Please rate this page
(literature.php)
(0/150)

BRENDA support

Literature summary for 3.4.22.B79 extracted from

  • Saisawang, C.; Saitornuang, S.; Sillapee, P.; Ubol, S.; Smith, D.; Ketterman, A.
    Chikungunya nsP2 protease is not a papain-like cysteine protease and the catalytic dyad cysteine is interchangeable with a proximal serine (2015), Sci. Rep., 5, 17125 .
    View publication on PubMedView publication on EuropePMC
Search for more literature for 3.4.22.B79

Cloned(Commentary)

Cloned (Comment) Organism
expression in Escherichia coli. Proteins are expressed as inclusion bodies under culturing at 37°C, and only partially expressed in a soluble form at 22°C Chikungunya virus

Protein Variants

Protein Variants Comment Organism
C478A significantly less activity than wild type for the 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp)substrate Chikungunya virus
additional information preparation of both full length and a truncated protease domain from the C-terminus of the nsP2 protein. The protease domain alone has different properties compared with the full length nsP2 protease Chikungunya virus
S482A significantly less activity than wild type for the 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp)substrate Chikungunya virus
W549A mutation involved in structural stability of the loop that contains the catalytic histidine residue Chikungunya virus
W549F mutation involved in structural stability of the loop that contains the catalytic histidine residue Chikungunya virus

Inhibitors

Inhibitors Comment Organism Structure
Cu2+ strong inhibition with substrate 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) Chikungunya virus
Mg2+ stimulation with substrates 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) and 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp), inhibition with substrate 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) Chikungunya virus
additional information not inhibitory: leupeptin, E-64. Poor inhibitors: chymostatin, PMSF; not inhibitory to full-length enzyme: NaCl up to 2 M, but 30-40% inhibition at 1 M with truncated protease Chikungunya virus
PMSF PMSF does not affect activity when using 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) as a substrate. With 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp), PMSF shows no effect on truncated protease and 30% inhibition of full-length NSP2 Chikungunya virus
Zn2+ strong inhibition with all substrates tested Chikungunya virus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.00033
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.00041
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.00041
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.00058
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.0007
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.00072
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.00076
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.00077
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.00086
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.00104
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.00109
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.00184
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.00269
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.00301
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.0045
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.0066
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.0074
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.009
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0093
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0099
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0364
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus

Metals/Ions

Metals/Ions Comment Organism Structure
Co2+ up to 2fold stimulation Chikungunya virus
Mg2+ stimulation with substrates 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) and 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp), inhibition with substrate 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) Chikungunya virus

Organism

Organism UniProt Comment Textmining
Chikungunya virus
-
-
-
Chikungunya virus ECSA E1:226V
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) + H2O
-
 Chikungunya virus 2-(N-methylamino)benzoyl-AGA + GIIETk(Dnp)
-
 ?
2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) + H2O
-
 Chikungunya virus ECSA E1:226V 2-(N-methylamino)benzoyl-AGA + GIIETk(Dnp)
-
 ?
2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) + H2O
-
 Chikungunya virus 2-(N-methylamino)benzoyl-AGC + GIIETk(Dnp)
-
 ?
2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) + H2O
-
 Chikungunya virus ECSA E1:226V 2-(N-methylamino)benzoyl-AGC + GIIETk(Dnp)
-
 ?
2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) + H2O
-
 Chikungunya virus 2-(N-methylamino)benzoyl-AGG + GIIETk(Dnp)
-
 ?
2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) + H2O
-
 Chikungunya virus ECSA E1:226V 2-(N-methylamino)benzoyl-AGG + GIIETk(Dnp)
-
 ?
additional information chikungunya nsP2 protease possesses different substrate specificity to the canonical alphavirus nsP2 polyprotein cleavage specificity Chikungunya virus ?
-
 ?
additional information chikungunya nsP2 protease possesses different substrate specificity to the canonical alphavirus nsP2 polyprotein cleavage specificity Chikungunya virus ECSA E1:226V ?
-
 ?

Temperature Stability [°C]

Temperature Stability Minimum [°C] Temperature Stability Maximum [°C] Comment Organism
42
-
 10 min, 50.4% residual activity for wild-type, 53% for mutant C478A, 50.4% for mutant S482A, 52.9% for mutant W549A, 54% for mutant W549F Chikungunya virus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.00052
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.00076
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.0011
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.0013
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.0013
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.0014
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.0015
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.0016
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.00164
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.0017
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.0019
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0021
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.0024
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.0027
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0029
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.0031
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.0032
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.0033
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.0034
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.0046
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.0075
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus

General Information

General Information Comment Organism
physiological function a serine that is close to the catalytical dyad is catalytically interchangeable with the dyad cysteine residue. The enzyme retains activity upon alanine replacement of either residue but a replacement of both cysteine and serine residues results in no detectable activity Chikungunya virus

kcat/KM [mM/s]

kcat/KM Value [1/mMs-1] kcat/KM Value Maximum [1/mMs-1] Substrate Comment Organism Structure
0.067
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.072
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.091
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.104
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.116
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549A, pH 7.5, 37°C Chikungunya virus
0.15
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus
0.157
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.164
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.177
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.179
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.179
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.187
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.21
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.224
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.242
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant C478A, pH 7.5, 37°C Chikungunya virus
0.29
-
 2-(N-methylamino)benzoyl-AGAGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.308
-
 2-(N-methylamino)benzoyl-AGGGIIETk(Dnp) mutant S482A, pH 7.5, 37°C Chikungunya virus
0.34
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) mutant W549F, pH 7.5, 37°C Chikungunya virus
0.35
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) full-length enzyme, pH 7.5, 37°C Chikungunya virus
0.361
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) wild-type enzyme, pH 7.5, 37°C Chikungunya virus
0.65
-
 2-(N-methylamino)benzoyl-AGCGIIETk(Dnp) truncated enzyme, pH 7.5, 37°C Chikungunya virus