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

DOI: 10.1038/s41467-017-02006-0

Authors: Matilde De Las Rivas (University of Zaragoza) , Erandi Lira-Navarrete (University of Zaragoza; University of Copenhagen) , Earnest James Paul Daniel (Case Western Reserve University) , Ismael Compañón (Universidad de La Rioja) , Helena Coelho (Universidade de Nova de Lisboa; CIC bioGUNE; University of the Basque Country) , Ana Diniz (Universidade de Nova de Lisboa) , Jesús Jiménez-Barbero (CIC bioGUNE; University of the Basque Country; Ikerbasque) , Jesús M. Peregrina (Universidad de La Rioja) , Henrik Clausen (University of Copenhagen) , Francisco Corzana (Universidad de La Rioja) , Filipa Marcelo (Universidade de Nova de Lisboa) , Gonzalo Jiménez-Osés (Universidad de La Rioja) , Thomas A. Gerken (Case Western Reserve University) , Ramon Hurtado-Guerrero (University of Zaragoza; Fundación ARAID)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 8

State: Published (Approved)
Published: December 2017
Diamond Proposal Number(s): 10121

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.

Journal Keywords: Enzyme mechanisms; Glycobiology; Kinetics

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I03-Macromolecular Crystallography

Added On: 08/01/2018 11:35

Documents:
s41467-017-02006-0.pdf

Discipline Tags:

Biochemistry Catalysis Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)