2. Academic Validation
  3. The interdomain flexible linker of the polypeptide GalNAc transferases dictates their long-range glycosylation preferences

The interdomain flexible linker of the polypeptide GalNAc transferases dictates their long-range glycosylation preferences

  • Nat Commun. 2017 Dec 5;8(1):1959. doi: 10.1038/s41467-017-02006-0.
Matilde de Las Rivas 1,  Erandi Lira-Navarrete 1 2,  Earnest James Paul Daniel 3,  Ismael Compañón 4,  Helena Coelho 5 6 7,  Ana Diniz 5,  Jesús Jiménez-Barbero 6 7 8,  Jesús M Peregrina 4,  Henrik Clausen 2,  Francisco Corzana 4,  Filipa Marcelo 5,  Gonzalo Jiménez-Osés 4,  Thomas A Gerken 3 9 10,  Ramon Hurtado-Guerrero 11 12
Affiliations

Affiliations

  • 1 BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, 50018, Spain.
  • 2 Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, School of Dentistry, University of Copenhagen, Copenhagen, DK-2200, Denmark.
  • 3 Department of Biochemistry, Case Western Reserve University, Cleveland, 44106, OH, USA.
  • 4 Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, E-26006, Logroño, Spain.
  • 5 UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, 2829-516, Portugal.
  • 6 CIC bioGUNE, Bizkaia Technology Park, Building 801A, 48170, Derio, Spain.
  • 7 Departament of Organic Chemistry II, Faculty of Science & Technology, University of the Basque Country, Leioa, Bizkaia, 48940, Spain.
  • 8 Ikerbasque, Basque Foundation for Science, Maria Diaz de Haro 13, 48009, Bilbao, Spain.
  • 9 Department of Pediatrics, Case Western Reserve University, Cleveland, 44106, OH, USA.
  • 10 Department of Chemistry, Case Western Reserve University, Cleveland, 44106, OH, USA.
  • 11 BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, 50018, Spain. rhurtado@bifi.es.
  • 12 Fundación ARAID, 50018, Zaragoza, Spain. rhurtado@bifi.es.
PMID: 29208955 DOI: 10.1038/s41467-017-02006-0
Abstract

The polypeptide GalNAc-transferases (GalNAc-Ts), that initiate mucin-type O-glycosylation, consist of a catalytic and a lectin domain connected by a flexible linker. In addition to recognizing polypeptide sequence, the GalNAc-Ts exhibit unique long-range N- and/or C-terminal prior glycosylation (GalNAc-O-Ser/Thr) preferences modulated by the lectin domain. Here we report studies on GalNAc-T4 that reveal the origins of its unique N-terminal long-range Glycopeptide specificity, which is the opposite of GalNAc-T2. The GalNAc-T4 structure bound to a monoglycopeptide shows that the GalNAc-binding site of its lectin domain is rotated relative to the homologous GalNAc-T2 structure, explaining their different long-range preferences. Kinetics and molecular dynamics simulations on several GalNAc-T2 flexible linker constructs show altered remote prior glycosylation preferences, confirming that the flexible linker dictates the rotation of the lectin domain, thus modulating the GalNAc-Ts' long-range preferences. This work for the first time provides the structural basis for the different remote prior glycosylation preferences of the GalNAc-Ts.

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