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Structural basis for the action of thermolysin.
D E Tronrud1, S L Roderick, B W Matthews
1Institute of Molecular Biology, Howard Hughes Medical Institute, University of Oregon, Eugene 97403.
Matrix (Stuttgart, Germany). Supplement
|January 1, 1992
Summary
High-resolution X-ray crystallography revealed how inhibitors bind to thermolysin. This structural insight aids in understanding enzyme mechanisms and designing targeted drugs for zinc proteases.
Area of Science:
- Biochemistry
- Structural Biology
- Enzymology
Background:
- Thermolysin is a zinc metalloprotease with a well-characterized active site.
- Understanding enzyme-inhibitor interactions is crucial for drug discovery.
- Previous studies provided limited stereochemical detail on thermolysin inhibition.
Purpose of the Study:
- To determine the binding modes of various inhibitors to thermolysin using X-ray crystallography.
- To elucidate the stereochemical mechanism of action for thermolysin and related zinc proteases.
- To rationalize observed differences in inhibitor binding kinetics.
Main Methods:
- High-resolution X-ray crystallography of thermolysin complexed with inhibitors.
- Analysis of inhibitor interactions including dipeptides, mercaptans, hydroxamates, N-carboxymethyl peptides, and phosphonamidates.
- Model building to support crystallographic analysis.
Main Results:
- Detailed modes of binding for diverse inhibitors were determined.
- Inhibitor interactions provided insights into enzyme-substrate interactions during catalysis.
- Phosphonamidates, proposed transition-state analogues, exhibited varied binding configurations.
- Dissimilar binding modes rationalized kinetic differences among chemically similar inhibitors.
Conclusions:
- The study provides a detailed stereochemical mechanism for thermolysin.
- Findings offer insights applicable to other zinc proteases like carboxypeptidase A and angiotensin converting enzyme.
- Three-dimensional enzyme structures facilitate rational drug design against specific targets.