[Study on the effect of some group-specific agents on clostridiopeptidase].
The interaction of clostridiopeptidase of Clostridium histolyticum with EDC, TNM and MA, the specific reagents for COOH-groups, tyrosine and lysine residues was studied. It was shown that at pH 6.0 EDC inactivates the enzyme. The inactivation process follows the pseudo-first order kinetics and is described by a second order rate constant equal to 1 M-1 min-1. The synthetic substrate does not prevent, in practical terms, the enzyme inactivation by EDC. At pH 8.0 TNM modifies about 19 tyrosine residues in the clostridiopeptidase molecule which is accompanied by marked inhibition of the enzyme activity (down to 70-90%). In this case, the inactivation process is not described by simple pseudo-first order kinetics but is characterized by two steps (fast and slow) with second order rate constants of approximately 14 and 3.5 M-1 min-1, respectively. The synthetic substrate partly prevents the inactivation of the enzyme by TNM and protects 11 tyrosine residues. The MA-induced incorporation of 13 +/- 3 maleyl groups into the clostridiopeptidase molecule in partially prevented by the synthetic substrate with protects the enzyme against inactivation. The data obtained suggest that lysine residues are seemingly included into the active center of clostridiopeptidase, whereas tyrosine residues provide for the maintenance of active conformation of the enzyme.
Balaevskaia
T O
TO
Solov'eva
N I
NI
Orekhovich
V N
VN
D003094:Collagen / Q000378:metabolism; D005663:Furans / Q000494:pharmacology*; D007700:Kinetics; D008299:Maleic Anhydrides / Q000494:pharmacology*; D008697:Methane / Q000031:analogs & derivatives*; D003012:Microbial Collagenase / Q000378:metabolism*; D013379:Substrate Specificity; D013774:Tetranitromethane / Q000494:pharmacology*; D014443:Tyrosine / Q000378:metabolism
D005663:Furans; D008299:Maleic Anhydrides; D014443:Tyrosine; D003094:Collagen; D003012:Microbial Collagenase; D013774:Tetranitromethane; D008697:Methane
Izuchenie deĭstviia nekotorykh gruppospetsificheskikh agentov na klostridiopeptidazu.