title: Sequences from the first fibronectin type III repeat of the neural cell adhesion molecule allow O-glycan polysialylation of an adhesion molecule chimera.
abstract: Polysialic acid is a developmentally regulated, anti-adhesive polymer that is added to N-glycans on the fifth immunoglobulin domain (Ig5) of the neural cell adhesion molecule (NCAM). We found that the first fibronectin type III repeat (FN1) of NCAM is required for the polysialylation of N-glycans on the adjacent Ig5 domain, and we proposed that the polysialyltransferases recognize specific sequences in FN1 to position themselves for Ig5 N-glycan polysialylation. Other studies identified a novel FN1 acidic surface patch and α-helix that play roles in NCAM polysialylation. Here, we characterize the contribution of two additional FN1 sequences, Pro(510)-Tyr(511)-Ser(512) (PYS) and Gln(516)-Val(517)-Gln(518) (QVQ). Replacing PYS or the acidic patch dramatically decreases the O-glycan polysialylation of a truncated NCAM protein, and replacing the α-helix or QVQ shifts polysialic acid to FN1 O-glycans in full-length NCAM. We also found that the FN1 domain of the olfactory cell adhesion molecule, a homologous but unpolysialylated protein, could partially replace NCAM FN1. Inserting Pro(510)-Tyr(511) eliminated N-glycan polysialylation and enhanced O-glycosylation of an NCAM- olfactory cell adhesion molecule chimera, and inserting other FN1 sequences unique to NCAM, predominantly the acidic patch, created a new polysialyltransferase recognition site. Taken together, our results highlight the role of the FN1 α-helix and QVQ sequences in N-glycan polysialylation and demonstrate that the acidic patch primarily functions in O-glycan polysialylation.
authors.affiliation: Department of Biochemistry and Molecular Genetics, College of Medicine, University of Illinois, Chicago, Illinois 60607, USA.
authors.forename: Deirdre A
authors.initials: DA
authors.lastname: Foley
authors.forename: Kristin G
authors.initials: KG
authors.lastname: Swartzentruber
authors.forename: Matthew G
authors.initials: MG
authors.lastname: Thompson
authors.forename: Shalu Shiv
authors.initials: SS
authors.lastname: Mendiratta
authors.forename: Karen J
authors.initials: KJ
authors.lastname: Colley
mesh_terms: D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D005353:Fibronectins; D006801:Humans; D019158:N-Acetylneuraminic Acid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D019006:Neural Cell Adhesion Molecules / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011134:Polysaccharides / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012799:Sialyltransferases / Q000235:genetics / Q000378:metabolism
chemical_list: D005353:Fibronectins; D019006:Neural Cell Adhesion Molecules; D011134:Polysaccharides; D011993:Recombinant Fusion Proteins; D012799:Sialyltransferases; D019158:N-Acetylneuraminic Acid
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