Dynamic interplay between catalytic and lectin domains of GalNAc-transferases modulates protein O-glycosylation

DOI: 10.1038/ncomms7937
PMID: 25939779

Authors: Erandi Lira-Navarrete (University of Zaragoza) , Matilde De Las Rivas (University of Zaragoza) , Ismael Compañón (Universidad de La Rioja) , María Carmen Pallarés (University of Zaragoza) , Yun Kong (University of Copenhagen) , Javier Iglesias-Fernández (King's College London) , Gonçalo J. L. Bernardes (University of Cambridge) , Jesús M. Peregrina (Universidad de La Rioja) , Carme Rovira (Universitat de Barcelona) , Pau Bernadó (INSERM U1054, CNRS UMR 5048, Université Montpellier 1 and 2) , Pierpaolo Bruscolini (University of Zaragoza) , Henrik Clausen (University of Copenhagen) , Anabel Lostao (University of Zaragoza) , Francisco Corzana (Universidad de La Rioja) , Ramon Hurtado-Guerrero (University of Zaragoza)
Co-authored by industrial partner: No

Type: Journal Paper
Journal: Nature Communications , VOL 6

State: Published (Approved)
Published: May 2015
Diamond Proposal Number(s): 8035 , 10121

Abstract: Protein O-glycosylation is controlled by polypeptide GalNAc-transferases (GalNAc-Ts) that uniquely feature both a catalytic and lectin domain. The underlying molecular basis of how the lectin domains of GalNAc-Ts contribute to glycopeptide specificity and catalysis remains unclear. Here we present the first crystal structures of complexes of GalNAc-T2 with glycopeptides that together with enhanced sampling molecular dynamics simulations demonstrate a cooperative mechanism by which the lectin domain enables free acceptor sites binding of glycopeptides into the catalytic domain. Atomic force microscopy and small-angle X-ray scattering experiments further reveal a dynamic conformational landscape of GalNAc-T2 and a prominent role of compact structures that are both required for efficient catalysis. Our model indicates that the activity profile of GalNAc-T2 is dictated by conformational heterogeneity and relies on a flexible linker located between the catalytic and the lectin domains. Our results also shed light on how GalNAc-Ts generate dense decoration of proteins with O-glycans.

Journal Keywords: Biological Sciences; Biophysics; Biochemistry

Diamond Keywords: Enzymes

Subject Areas: Biology and Bio-materials, Chemistry


Instruments: I03-Macromolecular Crystallography , I04-1-Macromolecular Crystallography (fixed wavelength)

Other Facilities: ALBA (Barcelona) and PETRA-III (DESY/Hamburg

Added On: 24/09/2015 13:17

Documents:
ncomms7937.pdf

Discipline Tags:

Non-Communicable Diseases Health & Wellbeing Cancer Biochemistry Chemistry Structural biology Life Sciences & Biotech

Technical Tags:

Diffraction Macromolecular Crystallography (MX)