title: The membrane attack mechanism of complement: photolabeling reveals insertion of terminal proteins into target membrane.
abstract: We have utilized a membrane-restricted, photoactivable glycolipid probe to investigate the protein-lipid interactions involved in complement (C) mediated lysis of a target membrane. The purified C proteins C5b-6, C7, C8, and C9 were added to artificial membrane vesicles containing the 14C-labeled photoreactive probe anchored in the outer monolayer of the membrane, and 6-carboxyfluorescein trapped in the lumen as an indicator for effective lysis. Irradiation of the membrane samples at different stages of functional complex assembly resulted in labeling of each of the 5 terminal C proteins, indicating that all 5 proteins become inserted into the hydrophobic milieu of the membrane during some stage of complex assembly. However, at the final stage of complex assembly, only C9 appeared to be labeled. Because we can demonstrate that the photoreactive probe has no strong affinity for C9 over the other terminal components (C5b-C8), the extensive change in labeling specificity during assembly is evidence for substantial changes in protein-lipid and possibly protein-protein interactions during formation of the C lesion.
mesh_terms: D002462:Cell Membrane / Q000276:immunology; D003182:Complement C5 / Q000378:metabolism; D003183:Complement C6 / Q000378:metabolism; D003184:Complement C7 / Q000378:metabolism; D003185:Complement C8 / Q000378:metabolism; D003186:Complement C9 / Q000378:metabolism; D003165:Complement System Proteins; D008433:Mathematics; D008563:Membrane Lipids / Q000378:metabolism*; D008565:Membrane Proteins / Q000378:metabolism*; D013696:Temperature; D013997:Time Factors
chemical_list: D003182:Complement C5; D003183:Complement C6; D003184:Complement C7; D003185:Complement C8; D003186:Complement C9; D008563:Membrane Lipids; D008565:Membrane Proteins; D003165:Complement System Proteins