title: Saposin D solubilizes anionic phospholipid-containing membranes.
abstract: Saposin (Sap) D is a late endosomal/lysosomal small protein, generated together with three other similar proteins, Sap A, B, and C, from the common precursor, prosaposin. Although the functions of saposins such as Sap B and C are well known (Sap B promotes the hydrolysis of sulfatides and Sap C that of glucosylceramide), neither the physiological function nor the mechanism of action of Sap D are yet fully understood. We previously found that a dramatic increase of Sap D superficial hydrophobicity, occurring at the low pH values characteristic of the late endosomal/lysosomal environment, triggers the interaction of the saposin with anionic phospholipid-containing vesicles. We have presently found that, upon lipid binding, Sap D solubilizes the membranes, as shown by the clearance of the vesicles turbidity. The results of gel filtration, density gradient centrifugation, and negative staining electron microscopy demonstrate that this effect is due to the transformation of large vesicles to smaller particles. The solubilizing effect of Sap D is highly dependent on pH, the lipid/saposin ratio, and the presence of anionic phospholipids; small variations in each of these conditions markedly influences the activity of Sap D. The present study documents the interaction of Sap D with membranes as a complex process. Anionic phospholipids attract Sap D from the medium; when the concentration of the saposin on the lipid surface reaches a critical value, the membrane breaks down into recombinant small particles enriched in anionic phospholipids. Our results suggest that the role played by Sap D is more general than promoting sphingolipid degradation, e.g. the saposin might also be a key mediator of the solubilization of intralysosomal/late endosomal anionic phospholipid-containing membranes.
authors.affiliation: Department of Metabolism and Pathological Biochemistry, Istituto Superiore Sanita', Viale Regina Elena 299, 00161 Rome, Italy.
authors.forename: F
authors.initials: F
authors.lastname: Ciaffoni
authors.forename: R
authors.initials: R
authors.lastname: Salvioli
authors.forename: M
authors.initials: M
authors.lastname: Tatti
authors.forename: G
authors.initials: G
authors.lastname: Arancia
authors.forename: P
authors.initials: P
authors.lastname: Crateri
authors.forename: A M
authors.initials: AM
authors.lastname: Vaccaro
mesh_terms: D000838:Anions; D002850:Chromatography, Gel; D006023:Glycoproteins / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008563:Membrane Lipids / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D049231:Saposins; D012995:Solubility
chemical_list: D000838:Anions; D006023:Glycoproteins; D008563:Membrane Lipids; D010743:Phospholipids; D049231:Saposins