title: rCNT2 extracellular cysteines, Cys(615) and Cys(649), are important for maturation and sorting to the plasma membrane.
abstract: rCNT2 is a purine-preferring concentrative nucleoside transporter implicated in the regulation of extracellular adenosine levels and purinergic signaling. This study addressed the analysis of the CNT2 C-terminus tail as a domain likely to be implicated in transporter sorting. The topological mapping of this segment revealed that Cys(615) and Cys(649) are important residues for the proper trafficking of CNT2 to the plasma membrane. The inhibition of protein disulfide isomerase (PDI) and ER glycosidase I and II impaired rCNT2 trafficking to the cell surface, similarly to Cys(615) and Cys(649) mutants. The present work suggests these two cysteine residues are relevant for the proper sorting of the transporter and its functional performance.
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Barcelona, Institute of Biomedicine (IBUB) and Oncology Program, National Biomedical Research Institute on Liver and Gastrointestinal Diseases (CIBER ehd), Instituto de Salud Carlos III, Barcelona, Spain.
authors.forename: Itziar
authors.initials: I
authors.lastname: Pinilla-Macua
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Barcelona, Institute of Biomedicine (IBUB) and Oncology Program, National Biomedical Research Institute on Liver and Gastrointestinal Diseases (CIBER ehd), Instituto de Salud Carlos III, Barcelona, Spain.
authors.forename: Ana
authors.initials: A
authors.lastname: Claudio-Montero
authors.affiliation: Department of Biochemistry and Molecular Biology, University of Barcelona, Institute of Biomedicine (IBUB) and Oncology Program, National Biomedical Research Institute on Liver and Gastrointestinal Diseases (CIBER ehd), Instituto de Salud Carlos III, Barcelona, Spain. Electronic address: mpastor@ub.edu.
authors.forename: Marçal
authors.initials: M
authors.lastname: Pastor-Anglada
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D016466:CHO Cells; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism*; D003412:Cricetulus; D003545:Cysteine; D004721:Endoplasmic Reticulum / Q000187:drug effects / Q000378:metabolism; D005110:Extracellular Space / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D026901:Membrane Transport Proteins / Q000737:chemistry* / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D021382:Protein Sorting Signals; D017434:Protein Structure, Tertiary; D021381:Protein Transport / Q000187:drug effects; D051381:Rats; D019163:Reducing Agents / Q000494:pharmacology
chemical_list: D026901:Membrane Transport Proteins; D021382:Protein Sorting Signals; D019163:Reducing Agents; C065171:cif nucleoside transporter; D003545:Cysteine
keywords: CNT2; Cysteine; Plasma membrane; Sorting