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Literature summary for 3.4.21.26 extracted from

  • Heinis, C.; Alessi, P.; Neri, D.
    Engineering a thermostable human prolyl endopeptidase for antibody-directed enzyme prodrug therapy (2004), Biochemistry, 43, 6293-6303.
    View publication on PubMed

Application

Application Comment Organism
medicine use of enzyme in antibody-directed enzyme prodrug therapy strategy, ADEPT. Coupling of enzyme mutant E289G to human antibody specific to the EDB domain of fibronectin, conjugate retains antigen binding and enzymatic activity Homo sapiens

Protein Variants

Protein Variants Comment Organism
E289G improved protein stability, 4fold longer half-life at 37┬░C compared to wild-type Homo sapiens

Organism

Organism UniProt Comment Textmining
Homo sapiens
-
recombinant enzyme
-

Purification (Commentary)

Purification (Comment) Organism
native enzyme expressed in Escherichia coli is rapidly thermally inactivated. Mutation E289G improves protein stability Homo sapiens

Source Tissue

Source Tissue Comment Organism Textmining
liver
-
Homo sapiens
-

Specific Activity [micromol/min/mg]

Specific Activity Minimum [┬Ámol/min/mg] Specific Activity Maximum [┬Ámol/min/mg] Comment Organism
0.096
-
pH 7.4, 22┬░C, substrate benzyloxycarbonyl-Gly-L-Pro-doxorubicin Homo sapiens
0.813
-
pH 7.4, 22┬░C, substrate benzyloxycarbonyl-Gly-L-Pro-melphalan Homo sapiens
4.066
-
pH 7.4, 22┬░C, substrate benzyloxycarbonyl-Gly-L-Pro-7-amido-4-methylcoumarin Homo sapiens

Substrates and Products (Substrate)

Substrates Comment Substrates Organism Products Comment (Products) Rev. Reac.
benzyloxycarbonyl-Gly-L-Pro-7-amido-4-methylcoumarin + H2O
-
Homo sapiens benzyloxycarbonyl-Gly-L-Pro + 7-amino-4-methylcoumarin
-
?
benzyloxycarbonyl-Gly-L-Pro-doxorubicin + H2O
-
Homo sapiens benzyloxycarbonyl-Gly-L-Pro + doxorubicin
-
?
benzyloxycarbonyl-Gly-L-Pro-melphalan + H2O
-
Homo sapiens benzyloxycarbonyl-Gly-L-Pro + melphalan
-
?

Temperature Stability [┬░C]

Temperature Stability Minimum [┬░C] Temperature Stability Maximum [┬░C] Comment Organism
34
-
wild-type, 50% loss of activy in 30 min Homo sapiens
37
-
wild-type, 20 min, 15% loss of activity in phosphate-buffered saline, 69% loss of activiy in human serum Homo sapiens
43
-
mutant E289G, 50% loss of activity in 30 min Homo sapiens