A propeptide-based biosensor for the selective detection of Vibrio cholerae using an environment-sensitive fluorophore.

Autor:	DeColli AA; Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA; Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA., Koolik IM; Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA; Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA., Seminara AB; Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA; Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Microbial Pathogenesis, Yale University School of Medicine, New Haven, CT 06536, USA., Hatzios SK; Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520, USA; Microbial Sciences Institute, Yale University, West Haven, CT 06516, USA; Department of Chemistry, Yale University, New Haven, CT 06520, USA. Electronic address: stavroula.hatzios@yale.edu.
Jazyk:	angličtina
Zdroj:	Cell chemical biology [Cell Chem Biol] 2022 Oct 20; Vol. 29 (10), pp. 1505-1516.e7.
DOI:	10.1016/j.chembiol.2022.09.003
Abstrakt:	Proteases are attractive targets for infectious disease diagnostics. Peptide-based sensors that are cleaved by pathogen proteases can provide a rapid readout of infection. However, identifying peptide substrates specific to a targeted pathogen is a significant challenge. Here, we demonstrate that a structured propeptide domain from a bacterial protease can be repurposed as a protease-activated biosensor of the cholera pathogen Vibrio cholerae. We found that the peptidase inhibitor I9 domain of the secreted V. cholerae protease IvaP is rapidly degraded by V. cholerae, but not by other intestinal bacteria. By conjugating the I9 domain to an environment-sensitive fluorophore, we developed a fluorescent probe that enables the species-specific detection of V. cholerae in mixed bacterial cultures without nonspecific cleavage by other bacteria or intestinal cells. Our findings demonstrate that the IvaP propeptide is sufficient to impart selectivity to a cleavage-based V. cholerae biosensor, suggesting I9 domains could potentially be harnessed for diagnostic applications. Competing Interests: Declaration of interests S.K.H. and A.A.D. are inventors on a patent application describing propeptide-based biosensors (PCT International Application No. PCT/US2021/039062). (Copyright © 2022 Elsevier Ltd. All rights reserved.)
Databáze:	MEDLINE
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