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Literature summary for 3.4.21.91 extracted from

  • Pastorino, B.A.; Peyrefitte, C.N.; Grandadam, M.; Thill, M.C.; Tolou, H.J.; Bessaud, M.
    Mutagenesis analysis of the NS2B determinants of the Alkhurma virus NS2B-NS3 protease activation (2006), J. Gen. Virol., 87, 3279-3283.
    View publication on PubMed
Search for more literature for 3.4.21.91

Cloned(Commentary)

Cloned (Comment) Organism
cloning of the NS3 protease domain (NS3 S1-E171), the active deltaNS2B–NS3pro complex (NS3pro domain fused with NS2B M48-A94 and L126-R131 regions), the deltaNS2B-NS3pro HDA complex (with S138A), mutated proteins and chimeric proteases with the NS2B cofactor from Langat virus and Dengue virus type 3 replacing the Alkhurma virus sequence. PCR products cloned into pET-DEST42 vector Alkhumra hemorrhagic fever virus

Protein Variants

Protein Variants Comment Organism
deltaV88 deletion mutant Alkhumra hemorrhagic fever virus
G68A substitution in the NS2B sequence, decreased activity compared with that of the wild-type enzyme Alkhumra hemorrhagic fever virus
G81A substitution in the NS2B sequence, decreased activity compared with that of the wild-type enzyme Alkhumra hemorrhagic fever virus
L73A substitution in the NS2B sequence, decreased activity compared with that of the wild-type enzyme Alkhumra hemorrhagic fever virus
Q77A substitution in the NS2B sequence, mutation has greater effects on substrate binding than on the reaction rate Alkhumra hemorrhagic fever virus
S138A inactive Alkhumra hemorrhagic fever virus
V88D reduced activity compared with that of the wild-type Alkhumra hemorrhagic fever virus
V88K reduced activity compared with that of the wild-type Alkhumra hemorrhagic fever virus
W60A substitution in the NS2B sequence, decreased activity compared with that of the wild-type enzyme Alkhumra hemorrhagic fever virus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.3
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant G68A Alkhumra hemorrhagic fever virus
0.3
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant G81A Alkhumra hemorrhagic fever virus
0.4
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant V88D Alkhumra hemorrhagic fever virus
0.4
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant W60A Alkhumra hemorrhagic fever virus
0.6
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide deltaNS2B-NS3pro Alkhumra hemorrhagic fever virus
1.7
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant L73A Alkhumra hemorrhagic fever virus
2.2
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant V88K Alkhumra hemorrhagic fever virus
2.6
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide chimeric proteases with the NS2B cofactor from Langat virus replacing the Alkhurma virus sequence Alkhumra hemorrhagic fever virus
15.9
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant Q77A Alkhumra hemorrhagic fever virus

Organism

Organism UniProt Comment Textmining
Alkhumra hemorrhagic fever virus
-
-
-

Purification (Commentary)

Purification (Comment) Organism
-
 Alkhumra hemorrhagic fever virus

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
N-alpha-benzoyl-L-arginine-p-nitroanilide + H2O no substrate-dependent precipitation for either Alkhurma virus mutants or chimeric proteins Alkhumra hemorrhagic fever virus N-alpha-benzoyl-L-arginine + p-nitroaniline
-
 ?

Synonyms

Synonyms Comment Organism
NS2B-NS3 protease
-
 Alkhumra hemorrhagic fever virus

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
0.0004
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant V88D Alkhumra hemorrhagic fever virus
0.0006
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant V88K Alkhumra hemorrhagic fever virus
0.0018
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant G81A Alkhumra hemorrhagic fever virus
0.0018
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant W60A Alkhumra hemorrhagic fever virus
0.0051
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide chimeric proteases with the NS2B cofactor from Langat virus replacing the Alkhurma virus sequence Alkhumra hemorrhagic fever virus
0.0058
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant G68A Alkhumra hemorrhagic fever virus
0.008
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant Q77A Alkhumra hemorrhagic fever virus
0.0089
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide mutant L73A Alkhumra hemorrhagic fever virus
0.015
-
 N-alpha-benzoyl-L-arginine-p-nitroanilide deltaNS2B-NS3pro Alkhumra hemorrhagic fever virus

Cofactor

Cofactor Comment Organism Structure
additional information chimeric proteases with the NS2B cofactor from Langat virus and Dengue virus type 3 replacing the Alkhurma virus sequence activate NS3pro Alkhumra hemorrhagic fever virus
NS2B protein tight complementarity of the protein sequences is necessary for NS2B-dependent activation of NS3. NS2B residues V88 and Q77 are critical for Alkhurma virus NS3 activation. Poor conservation of the NS2B cofactor sequence within members of the genus Flavivirus Alkhumra hemorrhagic fever virus