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Literature summary for 3.4.21.4 extracted from

  • Page, M.J.; Bleackley, M.R.; Wong, S.; MacGillivray, R.T.; Di Cera, E.
    Conversion of trypsin into a Na(+)-activated enzyme (2006), Biochemistry, 45, 2987-2993.
    View publication on PubMed
Search for more literature for 3.4.21.4

Activating Compound

Activating Compound Comment Organism Structure
Na+ wild-type, no activation, engineering of wild-type into a Na+-activated enzyme by replacing residues in the 170, 186, and 220 loops to those of coagulation factor Xa Streptomyces griseus

Protein Variants

Protein Variants Comment Organism
A171S/Y172S/G173S/N174F/E175I/E180M/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y contrary to wild-type, mutant may be activated by Na+, mutations in 170 loop increase susceptibility to Na+ further Streptomyces griseus
G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y twofold increase in KM-value Streptomyces griseus
Y172M/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y contrary to wild-type, mutant may be activated by Na+ Streptomyces griseus
Y172S/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y contrary to wild-type, mutant may be activated by Na+ Streptomyces griseus
Y172S/Y217E/P222K/Y224K/P225Y 30fold increase in KM-value, contrary to wild-type, mutant may be activated by Na+ Streptomyces griseus
Y217E/P222K/Y224K/P225Y twofold increase in KM-value Streptomyces griseus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
0.0089
-
 factor Xa wild-type, pH 8.0, 25°C Streptomyces griseus
0.0205
-
 factor Xa mutant Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
0.0222
-
 factor Xa mutant G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
0.285
-
 factor Xa mutant Y172S/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus

Organism

Organism UniProt Comment Textmining
Streptomyces griseus P00775
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
factor Xa + H2O
-
 Streptomyces griseus ?
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
3
-
 factor Xa mutant Y172S/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
9.25
-
 factor Xa mutant G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
77.7
-
 factor Xa mutant Y172S/Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
86.4
-
 factor Xa mutant Y217E/P222K/Y224K/P225Y, pH 8.0, 25°C Streptomyces griseus
124
-
 factor Xa wild-type, pH 8.0, 25°C Streptomyces griseus