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PMID: 29208955 Published · epublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

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

Nature communications ·Vol. 8 ·No. 1 ·2017-00-05 ·Pages 1959

de Las Rivas M, Lira-Navarrete E, Daniel EJP, Compañón I, Coelho H, Diniz A, Jiménez-Barbero J, Peregrina JM, Clausen H, Corzana F, Marcelo F, Jiménez-Osés G, Gerken TA, Hurtado-Guerrero R

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.

MeSH Terms
Amino Acid Sequence Binding Sites Catalytic Domain Chimera/genetics Cloning, Molecular Glycopeptides/chemistry,metabolism Glycosylation Humans Kinetics Lectins/chemistry,metabolism Models, Molecular Molecular Docking Simulation Molecular Dynamics Simulation Mutagenesis, Site-Directed N-Acetylgalactosaminyltransferases/chemistry,genetics,metabolism Peptides/chemistry,metabolism Protein Binding Protein Conformation Protein Interaction Domains and Motifs Substrate Specificity
Chemicals
Glycopeptides Lectins Peptides N-Acetylgalactosaminyltransferases polypeptide N-acetylgalactosaminyltransferase
Authors & Affiliations
14 authors, click to expand affiliations / ORCID
de Las Rivas Matilde
BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, 50018, Spain.
Lira-Navarrete Erandi
BIFI, University of Zaragoza, BIFI-IQFR (CSIC) Joint Unit, Mariano Esquillor s/n, Campus Rio Ebro, Edificio I+D, Zaragoza, 50018, Spain. | Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, School of Dentistry, University of Copenhagen, Copenhagen, DK-2200, Denmark.
Daniel Earnest James Paul
Department of Biochemistry, Case Western Reserve University, Cleveland, 44106, OH, USA.
Compañón Ismael
Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, E-26006, Logroño, Spain.
Coelho Helena
UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, 2829-516, Portugal. | CIC bioGUNE, Bizkaia Technology Park, Building 801A, 48170, Derio, Spain. | Departament of Organic Chemistry II, Faculty of Science & Technology, University of the Basque Country, Leioa, Bizkaia, 48940, Spain.
Diniz Ana
UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, 2829-516, Portugal.
Jiménez-Barbero Jesús
CIC bioGUNE, Bizkaia Technology Park, Building 801A, 48170, Derio, Spain. | Departament of Organic Chemistry II, Faculty of Science & Technology, University of the Basque Country, Leioa, Bizkaia, 48940, Spain. | Ikerbasque, Basque Foundation for Science, Maria Diaz de Haro 13, 48009, Bilbao, Spain.
Peregrina Jesús M
Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, E-26006, Logroño, Spain.
Clausen Henrik
Copenhagen Center for Glycomics, Department of Cellular and Molecular Medicine, School of Dentistry, University of Copenhagen, Copenhagen, DK-2200, Denmark.
Corzana Francisco ORCID
Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, E-26006, Logroño, Spain.
Marcelo Filipa
UCIBIO, REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade de Nova de Lisboa, Caparica, 2829-516, Portugal.
Jiménez-Osés Gonzalo
Departamento de Química, Universidad de La Rioja, Centro de Investigación en Síntesis Química, E-26006, Logroño, Spain.
Gerken Thomas A
Department of Biochemistry, Case Western Reserve University, Cleveland, 44106, OH, USA. | Department of Pediatrics, Case Western Reserve University, Cleveland, 44106, OH, USA. | Department of Chemistry, Case Western Reserve University, Cleveland, 44106, OH, USA.
Hurtado-Guerrero Ramon
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. | Fundación ARAID, 50018, Zaragoza, Spain. rhurtado@bifi.es.
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Article Info
Journal
Nature communications
Abbr.
Nat Commun
ISSN
2041-1723
DOI
10.1038/s41467-017-02006-0
Email
rhurtado@bifi.es
Published
2017-00-05
Epub
2017-00-05
Pages
1959
Language
English
Region
England
NLM ID
101528555
PMCID
PMC5716993
Subset
IM
Grants
NIGMS NIH HHS · R01 GM113534 · United States
NIGMS NIH HHS · U01 GM113534 · United States
External Links
PubMed source DOI: 10.1038/s41467-017-02006-0 PMC Full Text View journal details
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