2003
DOI: 10.1016/j.ab.2003.08.033
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Development of a fluorescence polarization assay to screen for inhibitors of the FtsZ/ZipA interaction

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Cited by 68 publications
(51 citation statements)
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“…However, only a 60% decrease in the fraction of bound Nd-ZipA was found upon the addition of CTZ-WT. These observations are in accordance with the different affinities of both peptides (15). As expected, the peptidemediated dissociation of the Nd*-ZipA⅐FtsZ polymer complex was similarly observed when the FtsZ polymer was formed in the presence of GTP and a GTP-regenerating system.…”
Section: Inhibition Of the Binding Of Nd-zipa To Ftsz Polymers By C-tsupporting
confidence: 75%
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“…However, only a 60% decrease in the fraction of bound Nd-ZipA was found upon the addition of CTZ-WT. These observations are in accordance with the different affinities of both peptides (15). As expected, the peptidemediated dissociation of the Nd*-ZipA⅐FtsZ polymer complex was similarly observed when the FtsZ polymer was formed in the presence of GTP and a GTP-regenerating system.…”
Section: Inhibition Of the Binding Of Nd-zipa To Ftsz Polymers By C-tsupporting
confidence: 75%
“…Fluorescence polarization competition assays using two different peptides (CTZ-WT and CTZ-MUT, as described under supplemental Methods) designed to mimic a segment of the C-terminal region of FtsZ demonstrated that they could bind with high affinity to the FtsZ-binding domain of ZipA (15). The binding affinity of CTZ-MUT to the C-terminal region of ZipA is an order of magnitude higher than the binding of CTZ-WT (15). We have used fluorescence correlation spectroscopy competition assays to measure the ability of these peptides to inhibit the binding of Nd-ZipA to FtsZ polymers.…”
Section: Inhibition Of the Binding Of Nd-zipa To Ftsz Polymers By C-tmentioning
confidence: 99%
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“…A library of 250 000 compounds has now been screened using this methodology to identify E. coli FtsZ/ZipA protein-protein interaction inhibitors. Among the screened compounds, 29 hits were found (Kenny et al, 2003). This approach offers a direct way to assay for small molecules in the absence of structural, or practically any other information about the target protein.…”
Section: Current Approaches To Increase the Chemical Diversity Of Biomentioning
confidence: 99%
“…The biochemical assays utilize purified recombinant or native proteins to recapitulate biological process in a well. Such assays assess enzymatic activities [24,25], receptor-ligand binding [26], or protein-protein interactions [27] in cell-free e nvironment. This approach has been a standard mainstream approach in the pharmaceutical industry for decades.…”
Section: Overview Of Biochemical and Cell-based Assaysmentioning
confidence: 99%