title: Preventing E-cadherin aberrant N-glycosylation at Asn-554 improves its critical function in gastric cancer.
abstract: E-cadherin is a central molecule in the process of gastric carcinogenesis and its posttranslational modifications by N-glycosylation have been described to induce a deleterious effect on cell adhesion associated with tumor cell invasion. However, the role that site-specific glycosylation of E-cadherin has in its defective function in gastric cancer cells needs to be determined. Using transgenic mice models and human clinical samples, we demonstrated that N-acetylglucosaminyltransferase V (GnT-V)-mediated glycosylation causes an abnormal pattern of E-cadherin expression in the gastric mucosa. In vitro models further indicated that, among the four potential N-glycosylation sites of E-cadherin, Asn-554 is the key site that is selectively modified with β1,6 GlcNAc-branched N-glycans catalyzed by GnT-V. This aberrant glycan modification on this specific asparagine site of E-cadherin was demonstrated to affect its critical functions in gastric cancer cells by affecting E-cadherin cellular localization, cis-dimer formation, molecular assembly and stability of the adherens junctions and cell-cell aggregation, which was further observed in human gastric carcinomas. Interestingly, manipulating this site-specific glycosylation, by preventing Asn-554 from receiving the deleterious branched structures, either by a mutation or by silencing GnT-V, resulted in a protective effect on E-cadherin, precluding its functional dysregulation and contributing to tumor suppression.
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Porto, Portugal.
authors.forename: S
authors.initials: S
authors.lastname: Carvalho
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.
authors.forename: T A
authors.initials: TA
authors.lastname: Catarino
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Porto, Portugal.
authors.forename: A M
authors.initials: AM
authors.lastname: Dias
authors.affiliation: Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, Wako, Saitama, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Kato
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.|Freie Universität Berlin, Institute of Chemistry and Biochemistry, Berlin, Germany.
authors.forename: A
authors.initials: A
authors.lastname: Almeida
authors.affiliation: Center for Molecular Biology, University of Heidelberg, Heidelberg, Germany.
authors.forename: B
authors.initials: B
authors.lastname: Hessling
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.
authors.forename: J
authors.initials: J
authors.lastname: Figueiredo
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Porto, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Gärtner
authors.affiliation: Institute for Systems and Robotics (ISR/IST), LARSyS, Instituto Superior Técnico, University of Lisbon, Lisboa, Portugal.
authors.forename: J M
authors.initials: JM
authors.lastname: Sanches
authors.affiliation: Center for Molecular Biology, University of Heidelberg, Heidelberg, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Ruppert
authors.affiliation: Molecular Biochemistry and Clinical Investigation, Osaka University Graduate School of Medicine, Osaka, Japan.
authors.forename: E
authors.initials: E
authors.lastname: Miyoshi
authors.affiliation: Complex Carbohydrate Research Center, Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.
authors.forename: M
authors.initials: M
authors.lastname: Pierce
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Medical Faculty, University of Porto, Porto, Portugal.|Department of Pathology, Hospital S. Joao, Porto, Portugal.
authors.forename: F
authors.initials: F
authors.lastname: Carneiro
authors.affiliation: Department of Biomolecular Systems, Max Planck Institute of Colloids and Interfaces, Potsdam, Germany.
authors.forename: D
authors.initials: D
authors.lastname: Kolarich
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Medical Faculty, University of Porto, Porto, Portugal.
authors.forename: R
authors.initials: R
authors.lastname: Seruca
authors.affiliation: Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, Wako, Saitama, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Yamaguchi
authors.affiliation: Systems Glycobiology Research Group, RIKEN-Max Planck Joint Research Center, RIKEN Global Research Cluster, Wako, Saitama, Japan.
authors.forename: N
authors.initials: N
authors.lastname: Taniguchi
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Porto, Portugal.|Medical Faculty, University of Porto, Porto, Portugal.
authors.forename: C A
authors.initials: CA
authors.lastname: Reis
authors.affiliation: Instituto de Investigação e Inovação em Saúde (Institute for Research and Innovation in Health), University of Porto, Portugal/Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP), Porto, Portugal.|Institute of Biomedical Sciences of Abel Salazar (ICBAS), University of Porto, Porto, Portugal.
authors.forename: S S
authors.initials: SS
authors.lastname: Pinho
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine / Q000235:genetics; D015820:Cadherins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D020134:Catalytic Domain / Q000235:genetics; D045744:Cell Line, Tumor; D004285:Dogs; D005753:Gastric Mucosa / Q000378:metabolism / Q000473:pathology; D006031:Glycosylation; D019073:HT29 Cells; D006801:Humans; D061985:Madin Darby Canine Kidney Cells; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017351:N-Acetylglucosaminyltransferases / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D013274:Stomach Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology
chemical_list: D015820:Cadherins; D001216:Asparagine; D017351:N-Acetylglucosaminyltransferases; C052254:alpha-1,6-mannosylglycoprotein beta 1,6-N-acetylglucosaminyltransferase