The sheep analogue of human CD59: purification and characterization of its complement inhibitory activity.
An inhibitor of the membrane attack complex of complement was isolated from the membranes of sheep erythrocytes. Fast protein liquid chromatography (FPLC) and affinity purification procedures for this sheep complement-inhibiting protein (SCIP) both yielded a pure protein with an apparent M(r) of 19,000 under reducing and non-reducing conditions. Incubation of the denatured protein with neuraminidase and Endo-F reduced the apparent M(r) to 18,000 and 15,000 respectively, while treatment with O-deglycosidase or phosphatidylinositol-specific phospholipase C (PIPLC) did not affect the apparent M(r). SCIP was detectable on erythrocytes and lymphocytes but not on platelets and could partially be removed by PIPLC treatment. Deglycosylation of the pure protein markedly reduced and PIPLC treatment abolished its activity. A monoclonal antibody (mAb) raised against sheep complement-inhibiting protein (SCIP) enhanced the susceptibility of sheep erythrocytes to lysis by homologous complement. SCIP inhibited complement after the stage of C5b-7 formation. Amino-terminal protein sequence was obtained and was shown to be similar to that of human CD59. All these features suggest that SCIP is the sheep equivalent of human CD59. Human CD59 has been reported to be species selective in that it inhibits complement from relatively few species. However, SCIP efficiently inhibited lysis of guinea-pig erythrocytes by complement from a wide range of species tested indicating that it is a potent and non-selective inhibitor of the membrane attack complex of complement (MAC).
van den Berg
C W
CW
Dept. of Medical Biochemistry, University of Wales College of Medicine, Cardiff, U.K.
Harrison
R A
RA
Morgan
B P
BP
D000595:Amino Acid Sequence; D000818:Animals; D015703:Antigens, CD / Q000302:isolation & purification* / Q000494:pharmacology; D015153:Blotting, Western; D018957:CD59 Antigens; D002853:Chromatography, Liquid; D015938:Complement Membrane Attack Complex / Q000037:antagonists & inhibitors*; D006461:Hemolysis / Q000276:immunology; D006801:Humans; D008562:Membrane Glycoproteins / Q000302:isolation & purification* / Q000494:pharmacology; D008969:Molecular Sequence Data; D043265:Phosphatidylinositol Diacylglycerol-Lyase; D054801:Phosphoinositide Phospholipase C; D010727:Phosphoric Diester Hydrolases; D012756:Sheep / Q000276:immunology*; D013045:Species Specificity
D015703:Antigens, CD; D018957:CD59 Antigens; D015938:Complement Membrane Attack Complex; D008562:Membrane Glycoproteins; D010727:Phosphoric Diester Hydrolases; D054801:Phosphoinositide Phospholipase C; D043265:Phosphatidylinositol Diacylglycerol-Lyase