Allosteric inhibitors of inducible nitric oxide synthase dimerization discovered via combinatorial chemistry
| Autor: | Kirk Mcmillan, Gonghua Pan, Michael M. Morrissey, Leslie J. Browne, Mark A. Polokoff, Baldwin John J, Ronald Vergona, Gary Phillips, Marc Adler, Michael Ohlmeyer, David D. Davey, John Parkinson, Cornell Mallari, Charles B. Glaser, Nolan H. Sigal, Shawn David Erickson, Keith A. Eagen, James J. Devlin, Tish A. Young, Ronald E. Dolle, Eric Blasko, Daniel Chelsky, Richard I. Feldman, Douglas S. Auld, Marc Whitlow |
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| Rok vydání: | 2000 |
| Předmět: |
Models
Molecular Oxygenase Allosteric regulation Nitric Oxide Synthase Type II Plasma protein binding Biology Endothelial NOS Nitric Oxide Nitric oxide Cell Line chemistry.chemical_compound Mice Allosteric Regulation Animals Combinatorial Chemistry Techniques Humans Binding site Enzyme Inhibitors Multidisciplinary Binding Sites Molecular Structure Biological Sciences Molecular biology Small molecule Rats Nitric oxide synthase Biochemistry chemistry biology.protein Nitric Oxide Synthase Dimerization Protein Binding |
| Zdroj: | Proceedings of the National Academy of Sciences of the United States of America. 97(4) |
| ISSN: | 0027-8424 |
| Popis: | Potent and selective inhibitors of inducible nitric oxide synthase (iNOS) (EC 1.14.13.39 ) were identified in an encoded combinatorial chemical library that blocked human iNOS dimerization, and thereby NO production. In a cell-based iNOS assay (A-172 astrocytoma cells) the inhibitors had low-nanomolar IC 50 values and thus were >1,000-fold more potent than the substrate-based direct iNOS inhibitors 1400W and N- methyl- l -arginine. Biochemical studies confirmed that inhibitors caused accumulation of iNOS monomers in mouse macrophage RAW 264.7 cells. High affinity ( K d ≈ 3 nM) of inhibitors for isolated iNOS monomers was confirmed by using a radioligand binding assay. Inhibitors were >1,000-fold selective for iNOS versus endothelial NOS dimerization in a cell-based assay. The crystal structure of inhibitor bound to the monomeric iNOS oxygenase domain revealed inhibitor–heme coordination and substantial perturbation of the substrate binding site and the dimerization interface, indicating that this small molecule acts by allosterically disrupting protein–protein interactions at the dimer interface. These results provide a mechanism-based approach to highly selective iNOS inhibition. Inhibitors were active in vivo , with ED 50 values of |
| Databáze: | OpenAIRE |
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