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The effect of chlorambucil upon lysozyme activity
A S Brecher1, E Rasmusson, C Riley
1Department of Chemistry, Bowling Green State University, OH 43403, USA.
Fundamental & Clinical Pharmacology
|February 23, 1999
Summary
Chicken egg white lysozyme inhibition was studied using diazoacetyl-DL-norleucine methyl ester (DANME) and chlorambucil. N,N',N"-triacetylchitotriose protected the enzyme from both inhibitors, suggesting specific active site interactions.
Area of Science:
- Biochemistry
- Enzymology
Background:
- Lysozyme is a crucial enzyme in the innate immune system.
- Understanding enzyme inhibition mechanisms is vital for drug development.
Purpose of the Study:
- To investigate the inhibitory effects of DANME and chlorambucil on chicken egg white lysozyme.
- To elucidate the protective role of N,N",N"-triacetylchitotriose [(NAG)3] against these inhibitors.
Main Methods:
- Enzyme kinetics assays were performed using M. lysodeikticus as a substrate.
- Lysozyme activity was measured over time in the presence of inhibitors and a protective agent.
- Inhibitor-enzyme and inhibitor-substrate interactions were analyzed.
Main Results:
- DANME inhibited lysozyme by 87% and chlorambucil by 93%.
- (NAG)3 provided 40% protection against DANME and 14% against chlorambucil.
- DANME's inhibition was partially uncompetitive, while chlorambucil's was mixed.
Conclusions:
- DANME likely inhibits lysozyme by esterifying aspartic acid 101.
- (NAG)3 protects the active site, preventing DANME-induced inactivation.
- Chlorambucil causes irreversible inhibition through broader alkylation and conformational changes.