Increased adhesion of erythrocytes to components of the extracellular matrix: isolation and characterization of a red blood cell lipid that binds thrombospondin and laminin.
Red blood cell (RBC) adhesion to the vascular endothelium is increased in several pathologic conditions, including sickle cell disease and malaria. However, RBC interactions with components of the subendothelial matrix are not well-characterized. Under in vitro flow conditions of 1 dyne/cm2, washed RBCs bound to the purified adhesive molecules thrombospondin (TSP) and laminin. Sickle RBCs had the greatest adhesion of all tested RBCs. The adhesion of sickle RBCs to immobilized TSP was inhibited by the anionic polysaccharides high molecular weight (MW) dextran sulfate and chondroitin sulfate A, but not other anionic polysaccharides of similar structure and/or charge density. These data were consistent with the RBC adhesive molecule being a sulfated glycolipid. Therefore, TSP-binding lipids from normal and sickle RBCs were isolated and characterized. The TSP-binding lipid was purified by alkaline methanolysis, anion exchange chromatography and preparative thin layer chromatography (TLC). A homogeneous band on TLC was identified using a specific overlay TSP-binding assay. TSP binding to the purified lipid was stable to bass and neuraminidase treatment, labile to acid treatment, and was inhibited by high MW dextran sulfate, similar to that seen with intact RBCs binding to immobilized TSP under conditions of flow. In addition, soluble laminin bound to the purified RBC lipid. This acidic TSP- and laminin-binding lipid(s) isolated from both sickle and normal RBC membranes may contribute to erythrocyte interactions with the subendothelial matrix, hereby participating in the pathogenesis of vaso-occlusive diseases.
Hillery
C A
CA
Department of Pediatrics, Medical College of Wisconsin, Milwaukee, USA.
Du
M C
MC
Montgomery
R R
RR
Scott
J P
JP
D000595:Amino Acid Sequence; D000755:Anemia, Sickle Cell / Q000097:blood; D002809:Chondroitin Sulfates / Q000494:pharmacology; D016264:Dextran Sulfate / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000378:metabolism*; D004912:Erythrocytes / Q000166:cytology*; D004913:Erythrocytes, Abnormal / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000378:metabolism*; D006017:Glycolipids / Q000378:metabolism*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007797:Laminin / Q000378:metabolism*; D008562:Membrane Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D008563:Membrane Lipids / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism; D019699:Thrombospondins
D016326:Extracellular Matrix Proteins; D006017:Glycolipids; D007797:Laminin; D008562:Membrane Glycoproteins; D008563:Membrane Lipids; D010446:Peptide Fragments; D019699:Thrombospondins; D002809:Chondroitin Sulfates; D016264:Dextran Sulfate