title: The aglycone of sulfogalactolipids can alter the sulfate ester substitution position required for hsc70 recognition.
abstract: 3'-Sulfogalactolipids(SGLs), sulfogalactosyl ceramide (SGC), and sulfogalactoglycerolipid (SGG) bind to the N-terminal ATPase-containing domain of members of the heat shock protein 70 family. We have probed this binding specificity using a series of synthetic positional sulfated or phosphorylated glycolipid analogues, containing either a long-chain bisalkyl hydrocarbon-2-(tetradecyl)hexadecane (B30) or C(18) ceramide (SGC(18)) backbone. By TLC overlay and receptor ELISA, recombinant hsc70 bound ceramide-based glycoconjugates having 3'- or 4'-sulfogalactose glycone moieties and the 4'-sulfogalactose positional isomer conjugated to B30. Hsc70 binding was significantly decreased to the 3'-sulfogalactose conjugated to the long-chain branched alkane. 3'-Sulfoglucose conjugated to B30 was not bound, nor were similarly conjugated di-, tri-, and tetra-sulfated or phosphorylated galactolipids. These results highlight the importance of the position, rather than the number of sulfate esters within the galactose ring. This binding selectivity was shared by the sea urchin hsp70-related sperm receptor. A 3'-SGC-based soluble inhibitor, in which the acyl chain was replaced with an adamantyl group, inhibited binding of hsc70 to both 3'- and 4'-SGC species with an IC(50) of 50 and 75 microM, respectively, indicating a shared sulfogalactose binding site. These studies demonstrate the highly specific nature of hsc70/SGL binding and show, for the first time, that the lipid aglycone can alter the substitution position requirement for glycolipid recognition.
authors.affiliation: Research Institute, Hospital for Sick Children, 555 University Avenue, Toronto, Ont., Canada M5G 1X8.
authors.forename: D
authors.initials: D
authors.lastname: Mamelak
authors.forename: M
authors.initials: M
authors.lastname: Mylvaganam
authors.forename: E
authors.initials: E
authors.lastname: Tanahashi
authors.forename: H
authors.initials: H
authors.lastname: Ito
authors.forename: H
authors.initials: H
authors.lastname: Ishida
authors.forename: M
authors.initials: M
authors.lastname: Kiso
authors.forename: C
authors.initials: C
authors.lastname: Lingwood
mesh_terms: D002554:Cerebrosides / Q000378:metabolism; D038983:Galactolipids; D005690:Galactose / Q000031:analogs & derivatives / Q000378:metabolism*; D006017:Glycolipids / Q000378:metabolism*; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000378:metabolism; D013379:Substrate Specificity; D013433:Sulfoglycosphingolipids / Q000378:metabolism; D013463:Sulfuric Acid Esters / Q000378:metabolism*
chemical_list: D002554:Cerebrosides; D038983:Galactolipids; D006017:Glycolipids; D050883:HSC70 Heat-Shock Proteins; D018840:HSP70 Heat-Shock Proteins; D011956:Receptors, Cell Surface; D013433:Sulfoglycosphingolipids; D013463:Sulfuric Acid Esters; C083015:cerebroside sulfate; C033639:glycolipid receptor; C011117:sulfoglycolipids; D005690:Galactose