Modulation of the Surface-Layer Protein of Clostridium difficile through Cwp84 Inhibition

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• 作者：Major D. Gooyit ; Kim D. Janda
• 刊名：ACS Infectious Diseases
• 出版年：2016
• 出版时间：July 8, 2016
• 年：2016
• 卷：2
• 期：7
• 页码：465-470
• 全文大小：363K
• ISSN：2373-8227

文摘

Cysteine protease Cwp84 is responsible for surface-layer processing in Clostridium difficile and was also shown to cleave several human extracellular matrix components in vitro. To enable the facile identification and characterization of Cwp84 inhibitors, we developed a fluorogenic 10-mer peptide based on the enzyme’s natural substrate SlpA that is amenable for use in FRET-based high-throughput screening. The design of substrate-mimetic inhibitors led to epoxysuccinate 8c, which displayed an inactivation efficiency (k_inact/K_I) of (4.7 ± 0.3) × 10⁴ M^–1 min^–1. Further evaluation of 8c demonstrated its ability to inhibit fibronectin cleavage and, more importantly, subvert surface-layer biogenesis in C. difficile.