| Assay Method Information | |
| | Luminescence Cell-Free Homogeneous Dose Response to Identify Inhibitors of Lux-S |
| Description: | Keywords: LuxS enzyme, AI-2 Assay Overview: The overall aim of the project is to identify inhibitors of the quorum sensing pathway in Vibrio harveyi. Previously, a primary assay was completed using a strain containing only the AI-2 quorum sensing system intact. The primary screen could potentially identify inhibitors of the AI-2 synthase LuxS, the AI-2 receptor LuxPQ, or other downstream elements of the pathway. Subsequent secondary assays and counterscreens such as the current assay will be used to determine the mode of action of compounds identified in the primary screen. Purified LuxS enzyme catalyzes the degradation of s-ribosylhomocysteine (SRH) to dihydroxypentadiene (DPD) and homocysteine. The homocysteine production is spectraphotometrically detected with Ellman's reagent. Expected Outcome: Inhibitors of LuxS will decrease the signal observed in the enzymatic assay. |
| Affinity data for this assay | |
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