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Nat Protoc. 2006;1(2):550-3.

A detergent-based assay for the detection of promiscuous inhibitors.

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  • 1Department of Pharmaceutical Chemistry, and Graduate Group in Chemistry and Chemical Biology, University of California-San Francisco, 1700 4th Street, California 94158-2330, USA.


At micromolar concentrations, many small molecules self-associate into colloidal aggregates that non-specifically inhibit enzymes and other proteins. Here we describe a protocol for identifying aggregate-based inhibitors and distinguishing them from small molecules that inhibit via specific mechanisms. As a convenient proxy for promiscuous, aggregate-based inhibition, we monitor inhibition of beta-lactamase in the absence and presence of detergent. Inhibition that is attenuated in the presence of detergent is characteristic of an aggregate-based mechanism. In the 96-well-format assay described here, about 200 molecules can be tested, in duplicate, per hour for detergent-dependent sensitivity. Furthermore, we also describe simple experiments that can offer additional confirmation of aggregate-based inhibition.

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