气候变化领域集成服务门户(CCPortal): A broadly active fucosyltransferase LmjFUT1 whose mitochondrial localization and activity are essential in parasitic Leishmania

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DOI	10.1073/pnas.2108963118
	A broadly active fucosyltransferase LmjFUT1 whose mitochondrial localization and activity are essential in parasitic Leishmania
	Guo H.; Damerow S.; Penha L.; Menzies S.; Polanco G.; Zegzouti H.; Ferguson M.A.J.; Beverley S.M.
发表日期	2021
ISSN	0027-8424
卷号	118 期号:33
英文摘要	Glycoconjugates play major roles in the infectious cycle of the trypanosomatid parasite Leishmania. While GDP-Fucose synthesis is essential, fucosylated glycoconjugates have not been reported in Leishmania major [H. Guo et al., J. Biol. Chem. 292, 10696-10708 (2017)]. Four predicted fucosyltransferases appear conventionally targeted to the secretory pathway; SCA1/2 play a role in side-chain modifications of lipophosphoglycan, while gene deletion studies here showed that FUT2 and SCAL were not essential. Unlike most eukaryotic glycosyltransferases, the predicted α 1-2 fucosyltransferase encoded by FUT1 localized to the mitochondrion. A quantitative “plasmid segregation” assay, expressing FUT1 from the multicopy episomal pXNG vector in a chromosomal null Δfut1− background, established that FUT1 is essential. Similarly, “plasmid shuffling” confirmed that both enzymatic activity and mitochondrial localization were required for viability, comparing import-blocked or catalytically inactive enzymes, respectively. Enzymatic assays of tagged proteins expressed in vivo or of purified recombinant FUT1 showed it had a broad fucosyltransferase activity including glycan and peptide substrates. Unexpectedly, a single rare Δfut1− segregant (Δfut1s) was obtained in rich media, which showed severe growth defects accompanied by mitochondrial dysfunction and loss, all of which were restored upon FUT1 reexpression. Thus, FUT1 along with the similar Trypanosoma brucei enzyme TbFUT1 [G. Bandini et al., bioRxiv, https://www.biorxiv.org/content/10.1101/726117v2 (2021)] joins the eukaryotic O-GlcNAc transferase isoform as one of the few glycosyltransferases acting within the mitochondrion. Trypanosomatid mitochondrial FUT1s may offer a facile system for probing mitochondrial glycosylation in a simple setting, and their essentiality for normal growth and mitochondrial function renders it an attractive target for chemotherapy of these serious human pathogens. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Chemotherapy; Fucose; Glycobiology; Glycosyltransferase; Trypanosomatid protozoan parasites
语种	英语
scopus关键词	ataxin 2; fucosyltransferase; fucosyltransferase LmjFUT1; lipophosphoglycan; stem cell antigen 1; unclassified drug; fucosyltransferase; galactoside 2-alpha-L-fucosyltransferase; protozoal protein; allele; Article; assay; cell survival; chemical modification; controlled study; disorders of mitochondrial functions; enzyme activity; enzyme glycosylation; FUT1 gene; gene; gene expression; growth disorder; Leishmania; mitochondrial membrane potential; nonhuman; parasite viability; plasmid segregation assay; protein expression; protein function; protein localization; secretory pathway; amino acid sequence; culture medium; enzymology; gene expression regulation; genetics; Leishmania major; metabolism; mitochondrion; mutation; physiology; plasmid; protein transport; Amino Acid Sequence; Culture Media; Fucosyltransferases; Gene Expression Regulation, Enzymologic; Leishmania major; Mitochondria; Mutation; Plasmids; Protein Transport; Protozoan Proteins
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/251073
作者单位	Department of Molecular Microbiology, Washington University, School of Medicine, St. Louis, MO 63110, United States; Division of Biological Chemistry and Drug Discovery, Wellcome Trust Biocentre, College of Life Science, University of Dundee, Scotland, Dundee, DD1 5EH, United Kingdom; R&D Department, Promega Corporation, Madison, WI 53711, United States
推荐引用方式 GB/T 7714	Guo H.,Damerow S.,Penha L.,et al. A broadly active fucosyltransferase LmjFUT1 whose mitochondrial localization and activity are essential in parasitic Leishmania[J],2021,118(33).
APA	Guo H..,Damerow S..,Penha L..,Menzies S..,Polanco G..,...&Beverley S.M..(2021).A broadly active fucosyltransferase LmjFUT1 whose mitochondrial localization and activity are essential in parasitic Leishmania.Proceedings of the National Academy of Sciences of the United States of America,118(33).
MLA	Guo H.,et al."A broadly active fucosyltransferase LmjFUT1 whose mitochondrial localization and activity are essential in parasitic Leishmania".Proceedings of the National Academy of Sciences of the United States of America 118.33(2021).