Immobilization Strategies for Functional Complement Convertase Assembly at Lipid Membrane Interfaces

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Immobilization Strategies for Functional Complement Convertase Assembly at Lipid Membrane Interfaces
Title:
Immobilization Strategies for Functional Complement Convertase Assembly at Lipid Membrane Interfaces
Other Titles:
Langmuir
Keywords:
Publication Date:
06 July 2017
Citation:
Langmuir 33, 29, 7332-7342
Abstract:
The self-assembly formation of complement convertases—essential biomacromolecular complexes that amplify innate immune responses—is triggered by protein adsorption. Herein, a supported lipid bilayer platform was utilized to investigate the effects of covalent and noncovalent tethering strategies on the self-assembly of alternative pathway C3 convertase components, starting with C3b protein adsorption followed bythe addition of factors B and D. Quartz crystal microbalance-dissipation (QCM-D) experiments measured the real-time kinetics of convertase assembly onto supported lipid bilayers. The results demonstrate that the nature of C3b immobilization onto supported lipid bilayers is a key factor governing convertase assembly. The covalent attachment of C3b to maleimide-functionalized supported lipid bilayers promoted the self-assembly of functional C3 convertase in the membrane-associated state and further enabled successful evaluation of a clinically relevant complement inhibitor, compstatin. By contrast, noncovalent attachment of C3b to negatively charged supported lipid bilayers also permitted C3b protein uptake, albeit membrane-associated C3b did not support convertase assembly in this case. Taken together, the findings in this work demonstrate that the attachment scheme for immobilizing C3b protein at lipid membrane interfaces is critical for downstream C3 convertase assembly, thereby offering guidance for the design and evaluation of membrane-associated biomacromolecular complexes.
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PublisherCopyrights
Funding Info:
Description:
ISSN:
0743-7463
1520-5827
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