Abstract
Broadly neutralizing antibodies such as 2F5 are directed against the membrane-proximal external region (MPER) of HIV-1 GP41 and recognize well-defined linear core sequences. These epitopes can be engrafted onto protein scaffolds to serve as immunogens with high structural fidelity. Although antibodies that bind to this core GP41 epitope can be elicited, they lack neutralizing activity. To understand this paradox, we used biophysical methods to investigate the binding of human 2F5 to the MPER in a membrane environment, where it resides in vivo. Recognition is stepwise, through a paratope more extensive than core binding site contacts alone, and dynamic rearrangement through an apparent scoop-like movement of heavy chain complementarity-determining region 3 (CDRH3) is essential for MPER extraction from the viral membrane. Core-epitope recognition on the virus requires the induction of conformational changes in both the MPER and the paratope. Hence, target neutralization through this lipid-embedded viral segment places stringent requirements on the plasticity of the antibody combining site.
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Acknowledgements
This work was supported by US National Institutes of Health (NIH) grants RO1AI84785 and U19AI91693 to E.L.R. and G.W., and a grant from the Gates Foundation, The Collaboration for AIDS Vaccine Discovery (CAVD) Program to E.L.R., G.W. and J.R.E. J.R.E. was also supported by NIH grant RO1-GM086507 and funding through a cooperative research agreement with the Waters Corporation. K.D.R. was supported by The Danish Council for Independent Research in Natural Sciences (FNU grant 09-063876). J.R.E. would like to thank T.E. Wales for expert technical assistance. L.S. was also supported by an US National High Magnetic Field Laboratory (NHMFL) User Collaboration Grants Program award. The NHMFL is funded by the US National Science Foundation through the Cooperative Agreement No. DMR-0654118, the State of Florida, and the US Department of Energy.
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Z.-Y.J.S. designed and conducted NMR experiments, analyzed the data and wrote the paper. G.W. directed the NMR effort and analyzed the NMR structure. A.F.F. contributed to the analysis of the CDRH3 loop. J.-H.W. contributed to the analysis of complex structure. X.S. and K.D.R. designed and carried out HX-MS experiments and contributed to the data analysis. J.R.E. contributed to the design and data analysis of HX-MS experiments and wrote the paper. L.S. conducted EPR experiments. S.M. prepared 2F5 and its variant Fab regions and carried out the labeling of MPER. Y.C. carried out cell transfection and FACS analysis. G.O., Y.Y. and P.D.K. provided 2F5 and mutant antibodies. M.K. designed and carried out SPR experiments, analyzed the data and wrote the paper. E.L.R. directed the project and wrote the paper.
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Kim, M., Sun, ZY., Rand, K. et al. Antibody mechanics on a membrane-bound HIV segment essential for GP41-targeted viral neutralization. Nat Struct Mol Biol 18, 1235–1243 (2011). https://doi.org/10.1038/nsmb.2154
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DOI: https://doi.org/10.1038/nsmb.2154
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