title: The cytoplasmic domain of syndecan-1 is required for cytoskeleton association but not detergent insolubility. Identification of essential cytoplasmic domain residues.
abstract: Syndecan-1 is a member of a gene family of multifunctional transmembrane heparan sulfate proteoglycans that bind a variety of extracellular ligands and possess highly conserved non-catalytic cytoplasmic domains. It has been shown that antibody-mediated clustering of syndecan-1 causes the proteoglycan to become associated with microfilaments and insoluble in non-ionic detergent. A series of truncation and point mutations of the syndecan-1 core protein was constructed to identify specific structural features that were required for these characteristics. The transmembrane domain but not the cytoplasmic domain was required for cell surface expression of syndecan-1. Deletion of the COOH-terminal 11 amino acids of the cytoplasmic domain had no effect, while deletion of an additional 12 amino acids abolished microfilament association. Mutation of a conserved tyrosine residue within the latter region also abolished microfilament association. In contrast, mutation of 2 tyrosine residues outside this region had no effect. Deletion of the entire cytoplasmic domain (except for a short stop-transfer sequence) did not affect insolubility of the proteoglycan in detergent. Analysis of a form of syndecan-1 that lacked glycosaminoglycan acceptor sites revealed that covalently attached glycosaminoglycans were not required for cell surface expression, microfilament association, or detergent insolubility. These results demonstrate that microfilament association is a function of a subregion within the cytoplasmic domain and suggest that insolubility in detergent is a function of the transmembrane domain.
authors.affiliation: Sigfried and Janet Weis Center for Research, Geisinger Clinic, Danville, Pennsylvania 17822, USA.
authors.forename: D J
authors.initials: DJ
authors.lastname: Carey
authors.forename: K M
authors.initials: KM
authors.lastname: Bendt
authors.forename: R C
authors.initials: RC
authors.lastname: Stahl
mesh_terms: D008841:Actin Cytoskeleton / Q000502:physiology / Q000648:ultrastructure; D000595:Amino Acid Sequence; D000818:Animals; D000831:Animals, Newborn; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D002478:Cells, Cultured; D003593:Cytoplasm / Q000378:metabolism; D003599:Cytoskeleton / Q000502:physiology* / Q000648:ultrastructure; D003902:Detergents; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D017354:Point Mutation; D011509:Proteoglycans / Q000737:chemistry / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012583:Schwann Cells / Q000378:metabolism* / Q000648:ultrastructure; D012584:Sciatic Nerve / Q000166:cytology / Q000378:metabolism; D017384:Sequence Deletion; D017386:Sequence Homology, Amino Acid; D012995:Solubility; D053668:Syndecan-1; D053667:Syndecans; D014162:Transfection; D014443:Tyrosine
chemical_list: D003902:Detergents; D008562:Membrane Glycoproteins; D011509:Proteoglycans; D011994:Recombinant Proteins; C508243:Sdc1 protein, rat; D053668:Syndecan-1; D053667:Syndecans; D014443:Tyrosine