2. Academic Validation
  3. Structures of DPAGT1 Explain Glycosylation Disease Mechanisms and Advance TB Antibiotic Design

Structures of DPAGT1 Explain Glycosylation Disease Mechanisms and Advance TB Antibiotic Design

  • Cell. 2018 Nov 1;175(4):1045-1058.e16. doi: 10.1016/j.cell.2018.10.037.
Yin Yao Dong 1,  Hua Wang 2,  Ashley C W Pike 1,  Stephen A Cochrane 3,  Sadra Hamedzadeh 2,  Filip J Wyszyński 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,  Wei-Min Liu 2,  Seung Seo Lee 2,  Takuya Machida 2,  Leanne Minall 2,  Shahid Mehmood 6,  Katsiaryna Belaya 7,  Wei-Wei Liu 7,  Amy Chu 1,  Leela Shrestha 1,  Shubhashish M M Mukhopadhyay 1,  Claire Strain-Damerell 1,  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 8,  Elisabeth P Carpenter 9
Affiliations

Affiliations

  • 1 Structural Genomics Consortium, University of Oxford, Oxford, OX3 7DQ, UK.
  • 2 Chemistry Research Laboratory, University of Oxford, Oxford, OX1 3TA, UK.
  • 3 Chemistry Research Laboratory, University of Oxford, Oxford, OX1 3TA, UK; 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.
  • 8 Chemistry Research Laboratory, University of Oxford, Oxford, OX1 3TA, UK. Electronic address: [email protected].
  • 9 Structural Genomics Consortium, University of Oxford, Oxford, OX3 7DQ, UK. Electronic address: [email protected].
PMID: 30388443 DOI: 10.1016/j.cell.2018.10.037
Abstract

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.

Keywords

DPAGT1; GPT; Protein N-glycosylation; congenital disorders of glycosylation; congenital myasthenic syndrome; tunicamycin.

Figures