title: Isolation and characterization of the eighth component of the bovine complement system.
abstract: OBJECTIVE
  To isolate and characterize the eighth component of the complement system (C8) in cattle.
  
  
  METHODS
  Fresh plasma obtained from beef cattle.
  
  
  METHODS
  Plasma samples were fractionated, using sequential precipitation and ion-exchange and gel-filtration chromatography, to yield C8. The protein was identified throughout the procedure on the basis of its hemolytic function. Electrophoresis in polyacrylamide gels was used to determine molecular weight and composition of polypeptide chains. Reconstitution of classical and alternative complement pathways was used to characterize the hemolytic function of bovine C8.
  
  
  RESULTS
  The bovine C8 protein consisted of a disulfide-bonded alpha-gamma heterodimer that was noncovalently associated with a beta chain. Apparent molecular weight of the alpha, beta, and gamma chains under reducing conditions were 66, 61, and 23 kd, respectively. In the classical pathway of activation, bovine C8 and the ninth component of the complement system (C9) had species incompatibility with human C8 and C9 on sheep erythrocyte target cells.
  
  
  CONCLUSIONS
  A simple 4-step fractionation procedure provided good yield of bovine C8 from plasma. The isolated protein was structurally comparable to C8 from other species. Purified bovine C8 may be useful in functional hemolytic assays to investigate the roles of complement-mediated lysis in the pathogenesis of inflammatory diseases and the killing of susceptible microorganisms.
authors.affiliation: Department of Microbiology and Immunology, Faculty of Health Sciences, Queen's University, Kingston, ON, Canada.
authors.forename: M
authors.initials: M
authors.lastname: Menger
authors.forename: R D
authors.initials: RD
authors.lastname: Sowery
authors.forename: W P
authors.initials: WP
authors.lastname: Aston
mesh_terms: D000818:Animals; D002417:Cattle / Q000276:immunology*; D003185:Complement C8 / Q000302:isolation & purification*; D004591:Electrophoresis, Polyacrylamide Gel / Q000662:veterinary; D006801:Humans; D008970:Molecular Weight; D011487:Protein Conformation
chemical_list: D003185:Complement C8