A detergent-based assay for the detection of promiscuous inhibitors
Top Cited Papers
- 27 June 2006
- journal article
- research article
- Published by Springer Nature in Nature Protocols
- Vol. 1 (2) , 550-553
- https://doi.org/10.1038/nprot.2006.77
Abstract
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 β-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.Keywords
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