Please wait a moment until all data is loaded. This message will disappear when all data is loaded.
Please wait a moment until the data is sorted. This message will disappear when the data is sorted.
E97N/Y99L
-
Km value for Pefachrome is 1.5fold higher than the Km-value of the wild-type enzyme. The KI-value for benzamidine is 92% of the wild-type value, the KI-value for [4-(6-chloro-naphthalene-2-sulfonyl)-piperazin-1-yl]-(3,4,5-6-tetrahydro-[2H-1,4']bipyridinyl-4yl)-methanone is 17% of the wild-type value, the KI-value for 2,7-bis(4-amidinobenzylidene)cycloheptan-1-one is 1.75fold higher than the wild-type value
E97N/Y99L/S190A
-
Km value for Pefachrome is 3.7fold higher than the Km-value of the wild-type enzyme. The KI-value for benzamidine is 1.3fold higher than wild-type value, the KI-value for [4-(6-chloro-naphthalene-2-sulfonyl)-piperazin-1-yl]-(3,4,5-6-tetrahydro-[2H-1,4']bipyridinyl-4yl)-methanone is 6.7% of the wild-type value, the KI-value for 2,7-bis(4-amidinobenzylidene)cycloheptan-1-one is 2.25fold higher than the wild-type value
S190A
-
Km value for Pefachrome is 1.9fold higher than the Km-value of the wild-type enzyme. The KI-value for benzamidine is 87% of the wild-type value, the KI-value for [4-(6-chloro-naphthalene-2-sulfonyl)-piperazin-1-yl]-(3,4,5-6-tetrahydro-[2H-1,4']bipyridinyl-4yl)-methanone is 60% of the wild-type value, the KI-value for 2,7-bis(4-amidinobenzylidene)cycloheptan-1-one is 75% of the wild-type value
R193F
-
the mutation does not affect the activation energy of the reaction, but significantly alters the preexponential term of the Arrhenius equation
R193G/G142A
-
reduced activity
R193G/G184A
-
reduced activity
R193G/G19A
-
reduced activity, completely resistant to chymotryptic degradation
R193Y
-
the mutation does not affect the activation energy of the reaction, but significantly alters the preexponential term of the Arrhenius equation
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
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
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
D189S/DELTA223
-
double mutant trypsinogen, the recombinant proteinase lacks trypsin-like activity but aquires a rather unique selectivity, it preferentially hydrolyses peptide bonds C-terminal to tyrosyl residues. This narrow specificity should be useful in peptide-analytical applications such as sequence-specific fragmentation of large proteins prior to sequencing
K188F
-
increase of Km for Arg and Lys containing substrates, cleavage of nearly 30 new peptide bonds in beta-casein compared to the wild-type enzyme
K188W
-
increase of Km for Arg and Lys containing substrates, 3-4fold decrease in turnover number
K188Y
-
increase of Km for Arg and Lys containing substrates, cleavage of nearly 30 new peptide bonds in beta-casein compared to the wild-type enzyme
N143H/E151H
-
the mutant enzyme with the engineered metal binding site hydrolyzes the peptide AGPYAHSS exclusively in presence of Ni2+ or Zn2+ with high levels of catalytic efficiency
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
G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
twofold increase in KM-value
L71R/S172A
-
site-directed mutagenesis, inactive active site mutant
L71Y
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type
L71Y/Q72R
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The active site mutant shows slightly lower activities toward type I collagen than the wild-type
Q72R
-
site-directed mutagenesis, the mutant shows a change in topological specificity and high hydrolytic activity toward type IV collagen. The mutant shows slightly lower activities toward type I collagen than the wild-type
S172A
-
site-directed mutagenesis, inactive active site mutant
T190A
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 47% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 55% of that of the wild-type enzyme
T190P
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 0.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.7% of that of the wild-type enzyme
T190S
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 18% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 9% of that of the wild-type enzyme
T190V
the ratio of turnover number to Km-value with tosyl-Gly-Pro-Arg-7-amido-4-methylcoumarin is 2.5% of that of the wild-type enzyme, the ratio of turnover number to Km-value with tosyl-Gly-Pro-Lys-7-amido-4-methylcoumarin is 0.6% of that of the wild-type enzyme
Y172M/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
contrary to wild-type, mutant may be activated by Na+
Y172S/G186K/G187Q/V188E/Y217E/P222K/Y224K/P225Y
contrary to wild-type, mutant may be activated by Na+
Y172S/Y217E/P222K/Y224K/P225Y
30fold increase in KM-value, contrary to wild-type, mutant may be activated by Na+
Y217E/P222K/Y224K/P225Y
twofold increase in KM-value
S172A
-
site-directed mutagenesis, inactive active site mutant
Y71L
-
site-directed mutagenesis, the mutant displays high activity toward type I collagen similar to the wild-type enzyme
Y71L/S172A
-
site-directed mutagenesis, inactive active site mutant
R193A
-
slightly reduced activity
R193A
-
the mutation does not affect the activation energy of the reaction, but significantly alters the preexponential term of the Arrhenius equation
R193G
-
completely resistant to chymotryptic degradation
R193G
-
the mutation does not affect the activation energy of the reaction, but significantly alters the preexponential term of the Arrhenius equation
additional information
-
attachment of O-carboxymethyl-poly-beta-cyclodextrin with molecular weight of 13000 Da to surface of enzyme. Resulting neoglycoenzyme retains high proteolytic and esterolytic activity, optimum temperature is increased by 10°C, enzyme is more resistant to thermal inactivation and to denaturation by sodiumdodecyl sulfate
additional information
Cambarus affinis
-
enzyme immobilized on Sepharose has higher thermal stability than the wild-type enzyme, enzyme immobilized on porous glass loaded with isothiocyanate propyl groups has the same thermal stability as the wild-type enzyme, enzyme immobilized on DEAE-Sephadex has lower thermal stability than the wild-type enzyme
additional information
-
activation of recombinant His-Patch ThioFusion enzyme by native enzyme
additional information
-
inhibition of trypsin expression by RNAi, enhances the survival of Leishmania mexicana in Lutzomyia longipalpis, overview. Trypsin 1 knockdown by dsRNA injection
additional information
-
loss of the Cys191-Cys220 disulfide has no effect on the stability of trypsin as measured by urea denaturation
additional information
-
the FX99 mutant is constructed using 19 replacements in wild type trypsin with residues present in FXa; i.e. N95T/G96K/T97E/insertionT98/insertionY99 in the 99-loop, A171S/Y172S/G173S/N174F/E175I/E180M in the 170-loop, DELTAP185/G186K/G187Q/V188E in the 186-loop, and Y217E/P222K/Y224K/P225Y in the 220-loop