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Structural and mechanistic characterization of bifunctional heparan sulfate N-deacetylase-N-sulfotransferase 1.

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العنوان: Structural and mechanistic characterization of bifunctional heparan sulfate N-deacetylase-N-sulfotransferase 1.
المؤلفون: Mycroft-West CJ; The Rosalind Franklin Institute, Harwell Science & Innovation Campus, OX11 0QX, Didcot, UK., Abdelkarim S; The Rosalind Franklin Institute, Harwell Science & Innovation Campus, OX11 0QX, Didcot, UK., Duyvesteyn HME; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, OX3 7BN, Oxford, UK., Gandhi NS; Department of Computer Science and Engineering, Manipal Institute of Technology, Manipal Academy of Higher Education, Manipal, 576104, Karnataka, India.; School of Chemistry and Physics, Queensland University of Technology, QLD 4000, Brisbane, Australia.; Centre for Genomics and Personalised Health, Queensland University of Technology, Brisbane, QLD 4059, Australia., Skidmore MA; Centre for Glycoscience Research and Training, Keele University, ST5 5BG, Newcastle-Under-Lyme, UK., Owens RJ; The Rosalind Franklin Institute, Harwell Science & Innovation Campus, OX11 0QX, Didcot, UK.; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, OX3 7BN, Oxford, UK., Wu L; The Rosalind Franklin Institute, Harwell Science & Innovation Campus, OX11 0QX, Didcot, UK. liang.wu@rfi.ac.uk.; Division of Structural Biology, Nuffield Department of Medicine, University of Oxford, The Wellcome Centre for Human Genetics, OX3 7BN, Oxford, UK. liang.wu@rfi.ac.uk.
المصدر: Nature communications [Nat Commun] 2024 Feb 13; Vol. 15 (1), pp. 1326. Date of Electronic Publication: 2024 Feb 13.
نوع المنشور: Journal Article
اللغة: English
بيانات الدورية: Publisher: Nature Pub. Group Country of Publication: England NLM ID: 101528555 Publication Model: Electronic Cited Medium: Internet ISSN: 2041-1723 (Electronic) Linking ISSN: 20411723 NLM ISO Abbreviation: Nat Commun Subsets: MEDLINE
أسماء مطبوعة: Original Publication: [London] : Nature Pub. Group
مواضيع طبية MeSH: Heparitin Sulfate*/metabolism , Sulfotransferases*/metabolism, Humans ; Cryoelectron Microscopy ; Catalytic Domain ; Extracellular Matrix/metabolism
مستخلص: Heparan sulfate (HS) polysaccharides are major constituents of the extracellular matrix, which are involved in myriad structural and signaling processes. Mature HS polysaccharides contain complex, non-templated patterns of sulfation and epimerization, which mediate interactions with diverse protein partners. Complex HS modifications form around initial clusters of glucosamine-N-sulfate (GlcNS) on nascent polysaccharide chains, but the mechanistic basis underpinning incorporation of GlcNS itself into HS remains unclear. Here, we determine cryo-electron microscopy structures of human N-deacetylase-N-sulfotransferase (NDST)1, the bifunctional enzyme primarily responsible for initial GlcNS modification of HS. Our structures reveal the architecture of both NDST1 deacetylase and sulfotransferase catalytic domains, alongside a non-catalytic N-terminal domain. The two catalytic domains of NDST1 adopt a distinct back-to-back topology that limits direct cooperativity. Binding analyses, aided by activity-modulating nanobodies, suggest that anchoring of the substrate at the sulfotransferase domain initiates the NDST1 catalytic cycle, providing a plausible mechanism for cooperativity despite spatial domain separation. Our data shed light on key determinants of NDST1 activity, and describe tools to probe NDST1 function in vitro and in vivo.
(© 2024. The Author(s).)
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معلومات مُعتمدة: United Kingdom WT_ Wellcome Trust
المشرفين على المادة: 9050-30-0 (Heparitin Sulfate)
EC 2.8.2.- (Sulfotransferases)
تواريخ الأحداث: Date Created: 20240213 Date Completed: 20240215 Latest Revision: 20240217
رمز التحديث: 20240218
مُعرف محوري في PubMed: PMC10864358
DOI: 10.1038/s41467-024-45419-4
PMID: 38351061
قاعدة البيانات: MEDLINE
الوصف
تدمد:2041-1723
DOI:10.1038/s41467-024-45419-4