Literature summary for 3.4.22.51 extracted from

Zhai, X.; Meek, T.D.
Catalytic mechanism of cruzain from Trypanosoma cruzi as determined from solvent kinetic isotope effects of steady-state and pre-steady-state kinetics (2018), Biochemistry, 57, 3176-3190 .

Search for more literature for 3.4.22.51

Protein Variants

Protein Variants	Comment	Organism
E208A	the mutant enzyme recognizes benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin less effectively than wild-type cruzain does, turnover of the substrate is comparable, suggesting that once bound, mutant cruzain processes benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarinat rates similar to that of the wild-type enzyme. Diminutions by 300fold and 200fold in the kcat/Km values of benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin and benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin, respectively, are observed for the E208A mutant cruzain compared to that of the wild type, suggesting that ion pairing between Glu208 and P2 Arg is essential for recognition of these substrates. This is also reflected in values of Km, which are increased for benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin (10fold) and benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin (4fold)	Trypanosoma cruzi

Inhibitors

Inhibitors	Comment	Organism	Structure
K11777	-	Trypanosoma cruzi

KM Value [mM]

KM Value [mM]	KM Value Maximum [mM]	Substrate	Comment	Organism
0.00054	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00056	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00061	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00062	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00068	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0007	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00077	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00087	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.6, 25°C, wild-type enzyme	Trypanosoma cruzi
0.00088	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0013	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0016	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
0.0019	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0025	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0029	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0036	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0037	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0041	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0043	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0046	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0052	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.0066	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.011	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.012	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.9, 25°C, wild-type enzyme	Trypanosoma cruzi
0.017	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.028	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
0.038	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.045	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
0.056	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
0.068	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
0.078	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.163	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi

Organism

Organism	UniProt	Comment	Textmining
Trypanosoma cruzi	P25779	-	-

Purification (Commentary)

Purification (Comment)	Organism
-	Trypanosoma cruzi

Substrates and Products (Substrate)

Substrates	Comment Substrates	Organism	Products	Comment (Products)	Rev.
benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin + H2O	-	Trypanosoma cruzi	benzyloxycarbonyl-Arg-Ala + 7-amino-4-methylcoumarin	-	?
benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin + H2O	-	Trypanosoma cruzi	benzyloxycarbonyl-Arg-Arg + 7-amino-4-methylcoumarin	-	?
benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin + H2O	-	Trypanosoma cruzi	benzyloxycarbonyl-Phe-Arg + 7-amino-4-methylcoumarin	-	?

Turnover Number [1/s]

Turnover Number Minimum [1/s]	Turnover Number Maximum [1/s]	Substrate	Comment	Organism
0.017	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
0.1	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
0.27	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
0.4	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
0.7	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
0.89	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
1.9	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
3.7	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.5, 25°C, wild-type enzyme	Trypanosoma cruzi
5.8	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
5.8	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.9, 25°C, wild-type enzyme	Trypanosoma cruzi
5.9	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
6.9	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
6.9	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
7.1	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
7.2	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
7.4	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
8	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
13	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
13	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
13	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
13.5	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
13.5	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
13.6	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.6, 25°C, wild-type enzyme	Trypanosoma cruzi
14	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
14	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
14	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
14.1	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
15	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
16	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
17	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
21	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi

kcat/KM [mM/s]

kcat/KM Value [1/mMs^-1]	kcat/KM Value Maximum [1/mMs^-1]	Substrate	Comment	Organism
0.2	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
1.2	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
5.9	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
6	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi
13	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
23	-	benzyloxycarbonyl-Arg-Ala-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
54	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
70	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
100	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
150	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
160	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
190	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
190	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
200	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
220	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 5.5, 25°C, wild-type enzyme	Trypanosoma cruzi
240	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
250	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 3.5, 25°C, wild-type enzyme	Trypanosoma cruzi
480	-	benzyloxycarbonyl-Arg-Arg-7-amido-4-methylcoumarin	pH 9.9, 25°C, wild-type enzyme	Trypanosoma cruzi
560	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.4, 25°C, wild-type enzyme	Trypanosoma cruzi
680	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 4.1, 25°C, wild-type enzyme	Trypanosoma cruzi
1100	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.0, 25°C, wild-type enzyme	Trypanosoma cruzi
1500	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.0, 25°C, wild-type enzyme	Trypanosoma cruzi
1500	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.5, 25°C, wild-type enzyme	Trypanosoma cruzi
1600	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 5.6, 25°C, wild-type enzyme	Trypanosoma cruzi
1900	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 6.5, 25°C, wild-type enzyme	Trypanosoma cruzi
2100	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.0, 25°C, wild-type enzyme	Trypanosoma cruzi
2600	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.0, 25°C, wild-type enzyme	Trypanosoma cruzi
2600	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 9.0, 25°C, wild-type enzyme	Trypanosoma cruzi
2700	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, wild-type enzyme	Trypanosoma cruzi
2700	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 8.5, 25°C, wild-type enzyme	Trypanosoma cruzi
13000	-	benzyloxycarbonyl-Phe-Arg-7-amido-4-methylcoumarin	pH 7.5, 25°C, mutant enzyme E208A	Trypanosoma cruzi