气候变化领域集成服务门户(CCPortal): A glycan FRET assay for detection and characterization of catalytic antibodies to the Cryptococcus neoformans capsule

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DOI	10.1073/PNAS.2016198118
	A glycan FRET assay for detection and characterization of catalytic antibodies to the Cryptococcus neoformans capsule
	Crawford C.J.; Wear M.P.; Smith D.F.Q.; d'Errico C.; McConnell S.A.; Casadevall A.; Oscarson S.
发表日期	2021
ISSN	00278424
卷号	118 期号:5
英文摘要	Classic antibody functions include opsonization, complement activation, and enhancement of cellular antimicrobial function. Antibodies can also have catalytic activity, although the contribution of catalysis to their biological functions has been more difficult to establish. With the ubiquity of catalytic antibodies against glycans virtually unknown, we sought to advance this knowledge. The use of a glycan microarray allowed epitope mapping of several monoclonal antibodies (mAbs) against the capsule of Cryptococcus neoformans. From this, we designed and synthesized two glycan-based FRET probes, which we used to discover antibodies with innate glycosidase activity and analyze their enzyme kinetics, including mAb 2H1, the most efficient identified to date. The validity of the FRET assay was confirmed by demonstrating that the mAbs mediate glycosidase activity on intact cryptococcal capsules, as observed by a reduction in capsule diameter. Furthermore, the mAb 18B7, a glycosidase hydrolase, resulted in the appearance of reducing ends in the capsule as labeled by a hydroxylamine-armed fluorescent (HAAF) probe. Finally, we demonstrate that exposing C. neoformans cells to catalytic antibodies results in changes in complement deposition and increased phagocytosis by macrophages, suggesting that the antiphagocytic properties of the capsule have been impaired. Our results raise questions over the ubiquity of antibodies with catalytic activity against glycans and establish the utility of glycan-based FRET and HAAF probes as tools for investigating this activity. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Chemical biology; Fungi; Glucuronoxylomannan; Glycans; Host-microbe interactions
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/181185
作者单位	Centre for Synthesis and Chemical Biology, University College Dublin, Dublin 4, Ireland; Department of Molecular Microbiology and Immunology, Johns Hopkins Bloomberg School of Public Health, Baltimore, MD 21205, United States
推荐引用方式 GB/T 7714	Crawford C.J.,Wear M.P.,Smith D.F.Q.,et al. A glycan FRET assay for detection and characterization of catalytic antibodies to the Cryptococcus neoformans capsule[J],2021,118(5).
APA	Crawford C.J..,Wear M.P..,Smith D.F.Q..,d'Errico C..,McConnell S.A..,...&Oscarson S..(2021).A glycan FRET assay for detection and characterization of catalytic antibodies to the Cryptococcus neoformans capsule.Proceedings of the National Academy of Sciences of the United States of America,118(5).
MLA	Crawford C.J.,et al."A glycan FRET assay for detection and characterization of catalytic antibodies to the Cryptococcus neoformans capsule".Proceedings of the National Academy of Sciences of the United States of America 118.5(2021).