The relationship between polymerization of complement component C9 and membrane channel formation.
C9 was studied with the objective to clarify the relationship between the process of C9 polymerization and membrane channel formation. Conditions that favor C9 polymerization include low ionic strength and calcium ion in the buffer. Moreover, polymerization is dependent on the concentration of C9. Calcium ion evokes about a threefold increase in the affinity constant for C9 self-association, and at 0 degrees C it imparts reversible amphiphilic properties in the molecule. These were discerned by measuring increases in the degree of reversible nonspecific binding of C9 to hydrophobic (tyramine-zymosan) and hydrophilic (arginyl-glutamyl-zymosan) supports as well as to erythrocytes. At 0 degrees C the hydrophilic-to-amphiphilic alteration of C9 is reversible, but upon incubation at 37 degrees C this transition is rendered permanent with the formation of poly(C9). A functional relationship between C9 polymerization and cytolysis was demonstrated by showing that polymerizing C9 can lyse reduced and alkylated erythrocytes. By studying comparative radiolabeling of tyrosine side chains within thrombin-nicked C9 and its polymerized form, it was demonstrated that upon polymerization the membrane-binding site of C9 becomes exposed. It is concluded that the process of circular polymerization of C9 causes a hydrophilic-to-amphiphilic transition that is required for membrane perforation and channel formation.
DiScipio
R G
RG
Research Institute of Scripps Clinic, Department of Immunology, La Jolla, CA 92037.
D000818:Animals; D002118:Calcium / Q000378:metabolism; D003186:Complement C9 / Q000378:metabolism*; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D003165:Complement System Proteins / Q000378:metabolism; D006461:Hemolysis; D006801:Humans; D066298:In Vitro Techniques; D007473:Ion Channels / Q000502:physiology*; D010743:Phospholipids / Q000378:metabolism; D011108:Polymers / Q000378:metabolism; D015032:Zinc / Q000494:pharmacology
D003186:Complement C9; D015938:Complement Membrane Attack Complex; D007473:Ion Channels; D010743:Phospholipids; D011108:Polymers; C042295:complement C5b-8 complex; D003165:Complement System Proteins; D015032:Zinc; D002118:Calcium