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Literature summary for 3.4.21.4 extracted from

  • Wang, E.C.W.; Hung, S.H.; Cahoon, M.; Hedstrom, L.
    The role of the Cys191-Cys220 disulfide bond in trypsin: new targets for engineering substrate specificity (1997), Protein Eng., 10, 405-411.
    View publication on PubMed
Search for more literature for 3.4.21.4

Cloned(Commentary)

Cloned (Comment) Organism
recombinant rat trypsinogen II is produced as an alpha-factor fusion protein in a Saccharomyces cerevisiae expression system from the pYT plasmid Rattus norvegicus

Protein Variants

Protein Variants Comment Organism
C191A mutation decreases the activity of trypsin by 20-200fold as measured by the ratio of turnover number to Km-value for the hydrolysis of amide substrates, ester hydrolysis is decreased by less than 10fold Rattus norvegicus
C191A/C220A the mutant enzyme binds bovine pancreatic trypsin inhibitor with the same affinity as trypsin, the affinity of benzamidine is decresed 10fold and the affinity of leupeptin is decreased 100fold Rattus norvegicus
C220A mutation decreases the activity of trypsin by 20-200fold as measured by the ratio of turnover number to Km-value for the hydrolysis of amide substrates, ester hydrolysis is decreased by less than 10fold Rattus norvegicus
additional information loss of the Cys191-Cys220 disulfide has no effect on the stability of trypsin as measured by urea denaturation Rattus norvegicus

KM Value [mM]

KM Value [mM] KM Value Maximum [mM] Substrate Comment Organism Structure
additional information
-
 additional information the mutations C191A and C220A decrease the activity of trypsin by 20-200fold as measured by the ratio of turnover number to Km-value for the hydrolysis of amide substrates, ester hydrolysis is decreased by less than 10fold Rattus norvegicus

Organism

Organism UniProt Comment Textmining
Rattus norvegicus
-
-
-

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzyloxycarbonyl-Arg-thiobenzyl ester + H2O
-
 Rattus norvegicus benzyloxycarbonyl-Arg + phenylmethanethiol
-
 ?
benzyloxycarbonyl-Lys-thiobenzyl ester + H2O
-
 Rattus norvegicus benzyloxycarbonyl-Lys + phenylmethanethiol
-
 ?
D-Val-Leu-Arg-4-methylcoumarin 7-amide + H2O
-
 Rattus norvegicus ?
-
 ?
D-Val-Leu-Lys-4-methylcoumarin 7-amide + H2O
-
 Rattus norvegicus ?
-
 ?
D-Val-Leu-Lys-thiobenzyl ester + H2O
-
 Rattus norvegicus ?
-
 ?
tosyl-Gly-Pro-Arg-4-methylcoumarin 7-amide + H2O
-
 Rattus norvegicus ?
-
 ?
tosyl-Gly-Pro-Lys-4-methylcoumarin 7-amide + H2O
-
 Rattus norvegicus ?
-
 ?

Turnover Number [1/s]

Turnover Number Minimum [1/s] Turnover Number Maximum [1/s] Substrate Comment Organism Structure
additional information
-
 additional information the mutations C191A and C220A decrease the activity of trypsin by 20-200fold as measured by the ratio of turnover number to Km-value for the hydrolysis of amide substrates, ester hydrolysis is decreased by less than 10fold Rattus norvegicus