Clusterin, the human apolipoprotein and complement inhibitor, binds to complement C7, C8 beta, and the b domain of C9.
Clusterin is a heterodimeric multifunctional protein expressed in a variety of tissues and cells. It forms high density lipid complexes in plasma and participates in the control of the lytic activity of the late complement complex (TCC, C5b-9). Together with vitronectin, clusterin binds to the nascent amphiphilic C5b-9 complex, rendering it water soluble and lytically inactive. To define the interactions that underlie the complement-inhibitory function of clusterin, we have examined the binding interactions between [125I]clusterin and the isolated components of the complex, C5b-6, C7, C8, and C9 and vitronectin. By using ligand blotting in the presence of Tween, specific binding of the labeled clusterin with C7, the beta-subunit of C8 and C9 was detected. Binding to C9 was competed by polymerized C9, but not by C8, C7, C6, and CD59, suggesting that the conformational change occurring during the hydrophilic-amphiphilic transition of C9 exposes the interaction site for clusterin. When thrombin-treated C9 was analyzed, clusterin was found to recognize the C9b fragment containing the hydrophobic membrane interaction segment. Both subunits of clusterin interact with C9 and are similarly potent in inhibiting C5b-9-mediated hemolysis and Zn+(+)-induced C9 polymerization. These results show that clusterin exerts its inhibitory effect by interacting with a structural motif common to C7, C8 alpha, and C9b.
Tschopp
J
J
Institute of Biochemistry, University of Lausanne, Epalinges, Switzerland.
Chonn
A
A
Hertig
S
S
French
L E
LE
D051152:Clusterin; D003184:Complement C7 / Q000378:metabolism*; D003185:Complement C8 / Q000378:metabolism*; D003186:Complement C9 / Q000378:metabolism*; D006023:Glycoproteins / Q000378:metabolism* / Q000494:pharmacology; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D066298:In Vitro Techniques; D018832:Molecular Chaperones; D010446:Peptide Fragments / Q000378:metabolism; D011485:Protein Binding
C495242:CLU protein, human; D051152:Clusterin; D003184:Complement C7; D003185:Complement C8; D003186:Complement C9; D006023:Glycoproteins; D018832:Molecular Chaperones; D010446:Peptide Fragments