Substrate activity screening (SAS): a general procedure for the preparation and screening of a fragment-based non-peptidic protease substrate library for inhibitor discovery
| Autor: | Andrew Patterson, Jonathan A. Ellman, Warren J. L. Wood |
|---|---|
| Rok vydání: | 2007 |
| Předmět: |
Fluorenes
Molecular Structure Drug discovery Drug design Computational biology Protein engineering Biology Plant biology Combinatorial chemistry General Biochemistry Genetics and Molecular Biology Substrate Specificity Structure-Activity Relationship Spectrometry Fluorescence Science research Plant science Coumarins Protease substrate Combinatorial Chemistry Techniques Protease Inhibitors Chromatography High Pressure Liquid |
| Zdroj: | Nature Protocols. 2:424-433 |
| ISSN: | 1750-2799 1754-2189 |
| DOI: | 10.1038/nprot.2007.28 |
| Popis: | Substrate activity screening (SAS) is a fragment-based method for the rapid development of novel substrates and their conversion into non-peptidic inhibitors of Cys and Ser proteases. The method consists of three steps: (i) a library of N-acyl aminocoumarins with diverse, low-molecular-weight N-acyl groups is screened to identify protease substrates using a simple fluorescence-based assay; (ii) the identified N-acyl aminocoumarin substrates are optimized by rapid analog synthesis and evaluation; and (iii) the optimized substrates are converted into inhibitors by direct replacement of the aminocoumarin with known mechanism-based pharmacophores. This protocol describes a general procedure for the solid-phase synthesis of a library of N-acyl aminocoumarin substrates and the screening procedure to identify weak binding substrates. |
| Databáze: | OpenAIRE |
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