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- W4366397674 abstract "Paracatalytic inducers are antagonists that shift the specificity of biological catalysts, resulting in non-native transformations. In this Chapter we describe methods to discover paracatalytic inducers of Hedgehog (Hh) protein autoprocessing. Native autoprocessing uses cholesterol as a substrate nucleophile to assist in cleaving an internal peptide bond within a precursor form of Hh. This unusual reaction is brought about by HhC, an enzymatic domain that resides within the C-terminal region of Hh precursor proteins. Recently, we reported paracatalytic inducers as a novel class of Hh autoprocessing antagonists. These small molecules bind HhC and tilt the substrate specificity away from cholesterol in favor of solvent water. The resulting cholesterol-independent autoproteolysis of the Hh precursor generates a non-native Hh side product with substantially reduced biological signaling activity. Protocols are provided for in vitro FRET-based and in-cell bioluminescence assays to discover and characterize paracatalytic inducers of Drosophila and human hedgehog protein autoprocessing, respectively." @default.
- W4366397674 created "2023-04-21" @default.
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- W4366397674 date "2023-01-01" @default.
- W4366397674 modified "2023-10-18" @default.
- W4366397674 title "Paracatalytic induction: Subverting specificity in hedgehog protein autoprocessing with small molecules" @default.
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- W4366397674 doi "https://doi.org/10.1016/bs.mie.2023.03.001" @default.
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