Abstract: A universal bacterial assay and kit for detecting and monitoring changes in concentration of a target analyte in solution. The assay relies on an engineered high affinity protein-based sensor with a detectable label that has high specificity to a target analyte, typically a ligand or metallated complex, and can be used to spectroscopically monitor interaction of the target analyte with a test bacterial strain. The sensors can be in the form of living, transport-deficient bacterial cells expressing the engineered high affinity protein, or in the form of purified proteins themselves. The system can further be used to screen candidate compounds for interference with the binding and uptake of such analytes by bacterial cells, which is useful in investigation of new classes of antibiotics.

Abstract: A cell-based assay for identifying a compound that inhibits iron transport in Gram-negative bacteria, engineered bacterial cells, and kits for conducting the same. The assay involves contacting a candidate compound with an engineered Gram-negative bacteria in the presence of iron for a sufficient period of time, exposing the reaction solution to an energy source to generate the detectable signal, and detecting changes in the detectable signal in the reaction solution over time. The engineered Gram-negative bacteria comprises an iron transport protein on its outer membrane that comprises an amino acid residue that has been engineered with a detectable label that generates a detectable signal. The changes in the detectable signal in the assay system over time correspond to the effect of the candidate compound on iron transport in the Gram-negative bacteria.

Abstract: A cell-based assay for identifying a compound that inhibits iron transport in Gram-negative bacteria, engineered bacterial cells, and kits for conducting the same. The assay involves contacting a candidate compound with an engineered Gram-negative bacteria in the presence of iron for a sufficient period of time, exposing the reaction solution to an energy source to generate the detectable signal, and detecting changes in the detectable signal in the reaction solution over time. The engineered Gram-negative bacteria comprises an iron transport protein on its outer membrane that comprises an amino acid residue that has been engineered with a detectable label that generates a detectable signal. The changes in the detectable signal in the assay system over time correspond to the effect of the candidate compound on iron transport in the Gram-negative bacteria.

Abstract: The present invention relates in general to spectroscopic methods for direct observation of membrane dynamics in living cells and their application to drug discovery methods. More particularly, the present invention relates to methods for observing iron transport activity in the outer membrane of living bacteria and more specifically to a ligand-specific opening of a gated-porin channel in the outer membrane of a living bacteria.

Abstract: The present invention relates in general to spectroscopic methods for direct observation of membrane dynamics in living cells and their application to drug discovery methods. More particularly, the present invention relates to methods for observing iron transport activity in the outer membrane of living bacteria and more specifically to a ligand-specific opening of a gated-porin channel in the outer membrane of a living bacteria.