In situ assembly of enzyme inhibitors using extended tethering.

来自 EBSCO

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作者：

DA Erlanson，JW Lam，C Wiesmann，TN Luong，RL Simmons，WL Delano，IC Choong，MT Burdett，WM Flanagan，D Lee

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摘要：

Cysteine aspartyl protease-3 (caspase-3) is a mediator of apoptosis and a therapeutic target for a wide range of diseases. Using a dynamic combinatorial technology, 'extended tethering', we identified unique nonpeptidic inhibitors for this enzyme. Extended tethering allowed the identification of ligands that bind to discrete regions of caspase-3 and also helped direct the assembly of these ligands into small-molecule inhibitors. We first designed a small-molecule 'extender' that irreversibly alkylates the cysteine residue of caspase-3 and also contains a thiol group. The modified protein was then screened against a library of disulfide-containing small-molecule fragments. Mass-spectrometry was used to identify ligands that bind noncovalently to the protein and that also form a disulfide linkage with the extender. Linking the selected fragments with binding elements from the extenders generates reversible, tight-binding molecules that are druglike and distinct from known inhibitors. One molecule derived from this approach inhibited apoptosis in cells.

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关键词：

particle accelerators enzyme inhibitors synchrotron radiation stanford linear accelerator center stanford synchrotron radiation laboratory

DOI：

10.1038/nbt786

被引量：

297

年份：

2003