title: Binding of human and rat CD59 to the terminal complement complexes.
abstract: CD59-antigen (protectin) is a widely distributed glycolipid-anchored inhibitor of complement lysis. CD59 interacts with complement components C8 and C9 during assembly of the membrane attack complex (MAC). To evaluate species specificity of these interactions we have in the present study examined cross-species binding of isolated human and rat CD59 to the terminal complement components C8 and C9. By using primarily soluble CD59 isolated from urine (CD59U) potentially non-specific binding interactions of the phospholipid portion of the membrane forms of CD59 could be avoided. Sucrose density gradient ultracentrifugation analysis showed that human CD59U bound to both human and rat C8 in the SC5b-8 complexes. Similar binding occurred when rat CD59U was used. The degree of binding did not significantly differ between the heterologous and homologous CD59-C8 combinations. C9 from both species inhibited the binding of CD59 to soluble SC5b-8. In ligand blotting analysis human and rat CD59U bound to human and rat C8 alpha gamma-subunit and C9. Binding of human and rat CD59U was stronger to human than rat C9. In plate binding assays the erythrocyte form of CD59 (CD59E) bound to both human and rat C8. Binding of CD59E to heterologous C9 was considerably weaker than to homologous C9. Our results imply that the reciprocal binding sites between C8 and CD59 and to a lesser degree between CD59 and C9 are conserved between human and rat. Interactions of CD59 with the terminal C components are thus species selective but not 'homologously restricted'.
authors.affiliation: Department of Bacteriology and Immunology, Haartman Institute, University of Helsinki, Finland.
authors.forename: T
authors.initials: T
authors.lastname: Lehto
authors.forename: B P
authors.initials: BP
authors.lastname: Morgan
authors.forename: S
authors.initials: S
authors.lastname: Meri
mesh_terms: D000818:Animals; D018957:CD59 Antigens / Q000378:metabolism*; D003185:Complement C8 / Q000378:metabolism*; D003186:Complement C9 / Q000378:metabolism*; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006023:Glycoproteins / Q000378:metabolism; D006801:Humans; D011869:Radioligand Assay; D051381:Rats; D013045:Species Specificity
chemical_list: D018957:CD59 Antigens; D003185:Complement C8; D003186:Complement C9; D015938:Complement Membrane Attack Complex; D006023:Glycoproteins; C047610:SC5b-9 protein complex; D003165:Complement System Proteins