부산대학교 도서관

Byline: Yin Yao Dong (1,15), Hua Wang (2,8,15), Ashley C.W. Pike (1,15), Stephen A. Cochrane (2,3,16), Sadra Hamedzadeh (2,16), Filip J. Wyszynski (2), Simon R. Bushell (1), Sylvain F. Royer (2), David A. Widdick (4), Andaleeb Sajid (5), Helena I. Boshoff (5), Yumi Park (5), Ricardo Lucas (2,9), Wei-Min Liu (2,10), Seung Seo Lee (2,11), Takuya Machida (2), Leanne Minall (2,12), Shahid Mehmood (6), Katsiaryna Belaya (7,13), Wei-Wei Liu (7), Amy Chu (1), Leela Shrestha (1), Shubhashish M.M. Mukhopadhyay (1), Claire Strain-Damerell (1,14), Rod Chalk (1), Nicola A. Burgess-Brown (1), Mervyn J. Bibb (4), Clifton E. Barry III (5), Carol V. Robinson (6), David Beeson (7), Benjamin G. Davis [Ben.Davis@chem.ox.ac.uk] (2,*), Elisabeth P. Carpenter [liz.carpenter@sgc.ox.ac.uk] (1,17,**) Keywords DPAGT1; GPT; Protein N-glycosylation; congenital myasthenic syndrome; congenital disorders of glycosylation; tunicamycin Highlights * Structures of DPAGT1 with UDP-GlcNAc and tunicamycin reveal mechanisms of catalysis * DPAGT1 mutations in patients with glycosylation disorders modulate DPAGT1 activity * Structures, kinetics and biosynthesis reveal role of lipid in tunicamycin * Lipid-altered, tunicamycin analogues give non-toxic antibiotics against TB Summary Protein N-glycosylation is a widespread post-translational modification. The first committed step in this process is catalysed by dolichyl-phosphate N-acetylglucosamine-phosphotransferase DPAGT1 (GPT/E.C. 2.7.8.15). Missense DPAGT1 variants cause congenital myasthenic syndrome and disorders of glycosylation. In addition, naturally-occurring bactericidal nucleoside analogues such as tunicamycin are toxic to eukaryotes due to DPAGT1 inhibition, preventing their clinical use. Our structures of DPAGT1 with the substrate UDP-GlcNAc and tunicamycin reveal substrate binding modes, suggest a mechanism of catalysis, provide an understanding of how mutations modulate activity (thus causing disease) and allow design of non-toxic 'lipid-altered' tunicamycins. The structure-tuned activity of these analogues against several bacterial targets allowed the design of potent antibiotics for Mycobacterium tuberculosis, enabling treatment in vitro, in cellulo and in vivo, providing a promising new class of antimicrobial drug. Author Affiliation: (1) Structural Genomics Consortium, University of Oxford, Oxford, OX3 7DQ, UK (2) Chemistry Research Laboratory, University of Oxford, Oxford, OX1 3TA, UK (3) School of Chemistry and Chemical Engineering, Queen's University, Belfast, UK (4) Department of Molecular Microbiology, John Innes Centre, Norwich, NR4 7UH, UK (5) Tuberculosis Research Section, Laboratory of Clinical Immunology and Microbiology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892, USA (6) Department of Chemistry, Oxford, OX1 3QZ, UK (7) Neurosciences Group, Nuffield Department of Clinical Neuroscience, Weatherall Institute of Molecular Medicine, University of Oxford, Oxford, OX3 9DS, UK * Corresponding author Article History: Received 21 March 2018; Revised 1 August 2018; Accepted 15 October 2018 (miscellaneous) Published: November 1, 2018 (footnote)8 Present address: The Francis Crick Institute, London, UK (footnote)9 Present address: Pharmacy, Seville University, Spain (footnote)10 Present address: Fu Jen Catholic University, Taiwan (footnote)11 Present address: Chemistry, University of Southampton, UK (footnote)12 Present address: Pfizer, Andover, MA, USA (footnote)13 Present address: Surgical Sciences, University of Oxford, UK (footnote)14 Present address: Diamond Light Source, Harwell, UK (footnote)15 These authors contributed equally (footnote)16 These authors contributed equally (footnote)17 Lead contact