title: N-glycosylation of the mammalian dipeptidyl aminopeptidase-like protein 10 (DPP10) regulates trafficking and interaction with Kv4 channels.
abstract: The dipeptidyl aminopeptidase-like protein 10 (DPP10) is a type II transmembrane protein homologue to the serine protease DPPIV/CD26 but enzymatically inactive. In the mammalian brain, DPP10 forms a complex with voltage-gated potassium channels of the Kv4 family, regulating their cell surface expression and biophysical properties. DPP10 is a glycoprotein containing eight predicted N-glycosylation sites in the extracellular domain. In this study we investigated the role of N-glycosylation on DPP10 trafficking and functional activity. Using site-directed mutagenesis (N to Q) we showed that N-glycosylation occured at six positions. Glycosylation at these specific residues was necessary for DPP10 trafficking to the plasma membrane as observed by flow cytometry. The surface expression levels of the substitutions N90Q, N119Q, N257Q and N342Q were reduced by more than 60%. Hence the interaction with the Kv4.3/KChIP2a channel complex was disrupted preventing the hastening effect of wild type DPP10 on current kinetics. Interestingly, N257 was crucial for this function and its substitution to glutamine completely blocked DPP10 sorting to the cell surface and prevented DPP10 dimerization. In summary, we demonstrated that glycosylation was necessary for both DPP10 trafficking to the cell surface and functional interaction with Kv4 channels.
authors.affiliation: Department of Health Sciences and Interdisciplinary Research Centre on Autoimmune Diseases (IRCAD), Università del Piemonte Orientale A. Avogadro, Via Solaroli 17, 28100 Novara, Italy. diego.cotella@med.unipmn.it
authors.forename: Diego
authors.initials: D
authors.lastname: Cotella
authors.forename: Susanne
authors.initials: S
authors.lastname: Radicke
authors.forename: Valentina
authors.initials: V
authors.lastname: Cipriani
authors.forename: Maria
authors.initials: M
authors.lastname: Cavaletto
authors.forename: Simone
authors.initials: S
authors.lastname: Merlin
authors.forename: Antonia
authors.initials: A
authors.lastname: Follenzi
authors.forename: Ursula
authors.initials: U
authors.lastname: Ravens
authors.forename: Erich
authors.initials: E
authors.lastname: Wettwer
authors.forename: Claudio
authors.initials: C
authors.lastname: Santoro
authors.forename: Daniele
authors.initials: D
authors.lastname: Sblattero
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001216:Asparagine / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D006031:Glycosylation; D006801:Humans; D047468:Immunoprecipitation; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D016296:Mutagenesis; D011485:Protein Binding; D021381:Protein Transport; D051672:Shal Potassium Channels / Q000378:metabolism*
chemical_list: D051672:Shal Potassium Channels; D001216:Asparagine; C476879:DPP10 protein, human; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases