title: Collagen fibril aggregation-inhibitor from sea cucumber dermis.
abstract: Collagen fibrils from the dermis of the sea cucumber Cucumaria frondosa are aggregated in vitro by the dermal glycoprotein stiparin (Trotter et al., 1996). Under physiological ionic conditions stiparin appears to be both necessary and sufficient to cause fibrils to aggregate (Trotter et al., 1997). We report here the initial biochemical and biophysical characterization of a sulfated glycoprotein from C. frondosa dermis that binds stiparin and inhibits its fibril-aggregating activity. This inhibitory glycoprotein, which has been named 'stiparin-inhibitor,' has the highest negative charge density of all the macromolecules extracted from the dermis. SDS-PAGE reveals three approximately 31-kDa bands that stain with alcian blue but not with Coomassie blue. Analytical ultracentrifugation indicates a native molecular weight of 62 kDa. Transmission electron microscopy of rotary-shadowed molecules shows curved rods about 22 nm long. The glycoprotein does not bind collagen fibrils, but does bind stiparin with a 1:1 stoichiometry. The binding of stiparin-inhibitor to stiparin prevents the binding of stiparin to collagen fibrils. The carbohydrate moiety produced by papain-digestion of the glycoprotein retains all of its inhibitory activity. The carbohydrate moiety of the inhibitor is dominated by galactose and sulfate.
authors.affiliation: Department of Cell Biology and Physiology, University of New Mexico School of Medicine, Albuquerque, NM 87131, USA. jtrotter@salud.unm.edu
authors.forename: J A
authors.initials: JA
authors.lastname: Trotter
authors.forename: G
authors.initials: G
authors.lastname: Lyons-Levy
authors.forename: K
authors.initials: K
authors.lastname: Chino
authors.forename: T J
authors.initials: TJ
authors.lastname: Koob
authors.forename: D R
authors.initials: DR
authors.lastname: Keene
authors.forename: M A
authors.initials: MA
authors.lastname: Atkinson
mesh_terms: D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001665:Binding Sites; D003094:Collagen / Q000737:chemistry* / Q000378:metabolism*; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D008854:Microscopy, Electron; D008970:Molecular Weight; D011485:Protein Binding; D012616:Sea Cucumbers / Q000378:metabolism*
chemical_list: D000596:Amino Acids; D006023:Glycoproteins; D003094:Collagen