title: Localization of glycosaminoglycan substitution sites on domain V of mouse perlecan.
abstract: Perlecan, the predominant basement membrane proteoglycan, has previously been shown to contain glycosaminoglycans attached at serine residues, numbers 65, 71, and 76, in domain I. However, the C-terminal domains IV and V of this molecule may also be substituted with glycosaminoglycan chains, but the exact substitution sites were not identified. The amino acid sequence of mouse perlecan reveals many ser-gly sequences in these domains that are possible sites for glycosaminoglycan substitution. We expressed recombinant domain IV and/or V of mouse perlecan in COS-7 cells and analyzed glycosaminoglycan substitution. Both heparan sulfate and chondroitin sulfate chains could be detected on recombinant domain V. One site, ser-gly-glu (serine residue 3593), toward the C-terminal region of domain V is a substitution site for heparan sulfate. When this sequence was absent, chondroitin/dermatan sulfate substitution was deleted, and the likely site for this galactosaminoglycan substitution was ser-gly-ala-gly (serine residue 3250) on domain V.
authors.affiliation: Department of Cell Biology, Cell Adhesion and Matrix Research Center, University of Alabama at Birmingham, Birmingham, Alabama, 35294-0019, USA.
authors.forename: P
authors.initials: P
authors.lastname: Tapanadechopone
authors.forename: J R
authors.initials: JR
authors.lastname: Hassell
authors.forename: B
authors.initials: B
authors.lastname: Rigatti
authors.forename: J R
authors.initials: JR
authors.lastname: Couchman
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites / Q000235:genetics; D019556:COS Cells; D017931:DNA Primers / Q000235:genetics; D006025:Glycosaminoglycans / Q000737:chemistry*; D006031:Glycosylation; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary / Q000235:genetics; D011509:Proteoglycans / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid; D014162:Transfection
chemical_list: D017931:DNA Primers; D006025:Glycosaminoglycans; D019812:Heparan Sulfate Proteoglycans; D006497:Heparan Sulfate; D011509:Proteoglycans; D011994:Recombinant Proteins; C071927:perlecan