On the cause and nature of C9-related heterogeneity of terminal complement complexes generated on target erythrocytes through the action of whole serum.
The binding of C8 and C9 from human serum to target erythrocytes was quantified, and the molecular stoichiometries of C9:C8 within terminal C5b-9(m) complexes were determined. Low doses of serum generated terminal complexes with mean C9:C8 ratios of 2 to 3:1, whereas complexes generated by highest serum doses harbored an average of six to eight C9/C8 molecules. From the collective biochemical and ultrastructural data, we concluded that heterogeneous populations of terminal complexes regularly form on target membranes; those containing high numbers of C9 molecules (greater than or equal to six to eight) exhibit the structure of the classical "lesion", whereas those containing low numbers of C9 do not exhibit this typical structure, although they probably still function as small pores. A major cause for this heterogeneity of the lesions derives from shortage of C9, which is naturally present in a 2 to 1 molar ratio relative to C8 in serum. Generation of terminal complexes harboring high numbers of C9 on erythrocyte membranes is possible in spite of this natural shortage because SC5b-9 does not form in the fluid phase to compete for C9 binding. If interrupted, the process of C9-C9 oligomerization cannot be recontinued, and "incomplete" C5b-9 complexes are unable to bind additional C9 upon reincubation with this component. The demonstrated heterogeneity of terminal complexes with respect to their C9 content may explain the functional heterogeneity of complement lesions observed previously by other investigators.
D000818:Animals; D001790:Blood Physiological Phenomena; D002499:Centrifugation, Density Gradient; D003185:Complement C8 / Q000032:analysis / Q000378:metabolism; D003186:Complement C9 / Q000032:analysis / Q000378:metabolism*; D015938:Complement Membrane Attack Complex; D003165:Complement System Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D003840:Deoxycholic Acid; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000378:metabolism* / Q000648:ultrastructure; D006461:Hemolysis; D006801:Humans; D011817:Rabbits; D011951:Receptors, Complement / Q000032:analysis
D003185:Complement C8; D003186:Complement C9; D015938:Complement Membrane Attack Complex; D011951:Receptors, Complement; D003840:Deoxycholic Acid; D003165:Complement System Proteins