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.

Details

Title
The interdomain flexible linker of the polypeptide GalNAc transferases dictates their long-range glycosylation preferences
Author
de las Rivas, Matilde 1 ; Lira-Navarrete, Erandi 2 ; Daniel, Earnest James Paul 3 ; Compañón, Ismael 4 ; Coelho, Helena 5 ; Diniz, Ana 6 ; Jiménez-Barbero, Jesús 7 ; Peregrina, Jesús M 4 ; Clausen, Henrik 8 ; Corzana, Francisco 4   VIAFID ORCID Logo  ; Filipa Marcelo 6 ; Jiménez-Osés, Gonzalo 4 ; Gerken, Thomas A 9 ; Hurtado-Guerrero, Ramon 10 

1  BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, Spain 
2  BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, Spain; Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, School of Dentistry, University of Copenhagen, Copenhagen, Denmark 
3  Department of Biochemistry, Case Western Reserve University, Cleveland, OH, USA 
4  Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, Logroño, Spain 
5  UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, Portugal; CIC bioGUNE, Bizkaia Technology Park, Derio, Spain; Departament of Organic Chemistry II, Faculty of Science & Technology, University of the Basque Country, Leioa, Bizkaia, Spain 
6  UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, Portugal 
7  CIC bioGUNE, Bizkaia Technology Park, Derio, Spain; Departament of Organic Chemistry II, Faculty of Science & Technology, University of the Basque Country, Leioa, Bizkaia, Spain; Ikerbasque, Basque Foundation for Science, Maria Diaz de Haro 13, Bilbao, Spain 
8  Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, School of Dentistry, University of Copenhagen, Copenhagen, Denmark 
9  Department of Biochemistry, Case Western Reserve University, Cleveland, OH, USA; Department of Pediatrics, Case Western Reserve University, Cleveland, OH, USA; Department of Chemistry, Case Western Reserve University, Cleveland, OH, USA 
10  BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, Spain; Fundación ARAID, Zaragoza, Spain 
Pages
1-11
Publication year
2017
Publication date
Dec 2017
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-017-02006-0
ProQuest document ID
1983426993
Copyright
© 2017. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.