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
  • 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. [email protected].
  • 12. Fundación ARAID, 50018, Zaragoza, Spain. [email protected].
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.