title: Comparison between complement and melittin hemolysis: anti-melittin antibodies inhibit complement lysis.
abstract: A comparison is made between the hemolytic actions of melittin and the ninth component of complement (C9). Melittin and C9 produce "pores" of similar effective radius in erythrocytes under standardized conditions, and their hemolytic action is suppressed by metal ions at similar concentrations, suggesting a common mechanism. Polyclonal anti-melittin immunoglobulin G (IgG) produced in rabbits retards hemolysis mediated by human C9 in a specific manner. Such antibodies react in several immunoassays with human and monkey C9 but not with C9 from lower animals, and no inhibition of lysis mediated by C9 molecules from these animals is observed. Thus, it is unlikely that anti-melittin IgG reacts with a structural element, such as an amphipathic helix, on human C9 since such structures are also predicted to exist in other C9 molecules. Human C9 and melittin block cross-reactivity in a dose-dependent manner, and anti-melittin IgG recognizes an epitope located between amino acid residues 245 and 390 of human C9 on "Western" blots. Comparison of the melittin and human C9 sequences indicates two regions of complete homology, a tetrapeptide at positions 292-295, and a pentapeptide at positions 527-531 in human C9, corresponding to residues 8-16 in melittin. Inhibition of hemolysis is not caused by blocking of C9 binding to the C5b-8 complex; rather the antibody must dissociate from the bound C9 before lysis ensues, indicating that it interferes with a postbinding event. It is proposed that anti-melittin binds to a conformational epitope on native, folded human C9 and thereby retards unfolding of the molecule, which is required for membrane insertion and hemolysis.
authors: Department of Comparative and Experimental Pathology, University of Florida, Gainesville 32610.
authors: R O
authors: RO
authors: Laine
authors: B P
authors: BP
authors: Morgan
authors: A F
authors: AF
authors: Esser
mesh_terms: D000906:Antibodies; D001514:Bee Venoms / Q000378:metabolism*; D003185:Complement C8 / Q000378:metabolism; D003186:Complement C9 / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism*; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000032:analysis; D006801:Humans; D007163:Immunosorbent Techniques; D007700:Kinetics; D008555:Melitten / Q000276:immunology / Q000378:metabolism*
chemical_list: D000906:Antibodies; D001514:Bee Venoms; D003185:Complement C8; D003186:Complement C9; D000939:Epitopes; D008555:Melitten; D003165:Complement System Proteins