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DOI	10.1073/pnas.2008815118
	Small-molecule inhibitors for the Prp8 intein as antifungal agents
	Li Z.; Tharappel A.M.; Xu J.; Lang Y.; Green C.M.; Zhang J.; Lin Q.; Chaturvedi S.; Zhou J.; Belfort M.; Li H.
发表日期	2021
ISSN	00278424
卷号	118 期号:2
英文摘要	Self-splicing proteins, called inteins, are present in many human pathogens, including the emerging fungal threats Cryptococcus neoformans (Cne) and Cryptococcus gattii (Cga), the causative agents of cryptococcosis. Inhibition of protein splicing in Cryptococcus sp. interferes with activity of the only intein-containing protein, Prp8, an essential intron splicing factor. Here, we screened a small-molecule library to find addititonal, potent inhibitors of the Cne Prp8 intein using a split-GFP splicing assay. This revealed the compound 6G-318S, with IC50 values in the low micromolar range in the split-GFP assay and in a complementary split-luciferase system. A fluoride derivative of the compound 6G-318S displayed improved cytotoxicity in human lung carcinoma cells, although there was a slight reduction in the inhibition of splicing. 6G-318S and its derivative inhibited splicing of the Cne Prp8 intein in vivo in Escherichia coli and in C. neoformans. Moreover, the compounds repressed growth of WT C. neoformans and C. gattii. In contrast, the inhibitors were less potent at inhibiting growth of the inteinless Candida albicans. Drug resistance was observed when the Prp8 intein was overexpressed in C. neoformans, indicating specificity of this molecule toward the target. No off-target activity was observed, such as inhibition of serine/cysteine proteases. The inhibitors bound covalently to the Prp8 intein and binding was reduced when the active-site residue Cys1 was mutated. 6G-318S showed a synergistic effect with amphotericin B and additive to indifferent effects with a few other clinically used antimycotics. Overall, the identification of these small-molecule intein-splicing inhibitors opens up prospects for a new class of antifungals. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Antifungal; Cryptococcus; Protein splicing; Prp8 intein; Small-molecule inhibitor
语种	英语
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/181047
作者单位	Wadsworth Center, New York State Department of Health, Albany, NY 12208, United States; Chemical Biology Program, Department of Pharmacology and Toxicology, University of Texas, Medical Branch, Galveston, TX 77555, United States; Department of Biological Sciences, RNA Institute, University at Albany, Albany, NY 12222, United States; RNA Epitranscriptomics and Proteomics Resource, University at Albany, Albany, NY 12222, United States; Department of Biomedical Sciences, School of Public Health, University at Albany, Albany, NY 12201-0509, United States; Department of Pharmacology and Toxicology, College of Pharmacy, University of Arizona, Tucson, AZ 85721, United States
推荐引用方式 GB/T 7714	Li Z.,Tharappel A.M.,Xu J.,et al. Small-molecule inhibitors for the Prp8 intein as antifungal agents[J],2021,118(2).
APA	Li Z..,Tharappel A.M..,Xu J..,Lang Y..,Green C.M..,...&Li H..(2021).Small-molecule inhibitors for the Prp8 intein as antifungal agents.Proceedings of the National Academy of Sciences of the United States of America,118(2).
MLA	Li Z.,et al."Small-molecule inhibitors for the Prp8 intein as antifungal agents".Proceedings of the National Academy of Sciences of the United States of America 118.2(2021).