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Role of C8 binding protein in homologous species restriction of C-mediated lysis: the C8bp interacts with. cap alpha. -. gamma. subunit of C8, and inhibits C9 polymerization

Conference · · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)
OSTI ID:5385484

The lysis of human (hu) erythrocytes (E) by C5b-9 is inefficient when hu C8/C9 are used to lyse EC5b-7. In the classical pathway activation, the ratio of C9 bound/inserted in hu E lysed with hu C is much lower when compared to sheep E. Recently, the authors have identified a protein from hu E that binds to hu C8. This C8 binding protein (C8bp) isolated from E inhibited lysis of chicken EC5b-7 by hu C8/C9, but not rabbit C8/C9. When hu C8/C9 were added to hu EC5b-7 with C8bp, poly C9 formation was inhibited. In the present study, the interaction of C8bp with C8 was further explored with /sup 125/I-labeled C8bp. Hu C8 was separated by SDS-PAGE under reducing or non-reducing conditions, transferred onto nitrocellulose paper by Western-blot. The blots were reacted with /sup 125/I-C8bp. Autoradiographic analysis showed the C8bp bands corresponding to the ..cap alpha..-..gamma.. subunit in a non-reduced gel and the ..gamma..-chain in a reduced gel, indicating direct interaction between C8bp and ..gamma..-chain of C8. Since the ..cap alpha gamma..-subunit does not bind to C5b-7, the authors' observation also explains the failure of C8bp to inhibit C8 binding to EC5b-7 shown previously. Inhibition of C5b-8 induced C9 polymerization by C8bp also suggests a direct effect of C8bp on C9 interaction with EC5b-8.

Research Organization:
Univ. of Heidelberg, West Germany
OSTI ID:
5385484
Report Number(s):
CONF-8604222-
Journal Information:
Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States), Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Vol. 45:3; ISSN FEPRA
Country of Publication:
United States
Language:
English