Structure-based design of a protein immunogen that displays an HIV-1 gp41 neutralizing epitope |
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Authors: | Stanfield Robyn L Julien Jean-Philippe Pejchal Robert Gach Johannes S Zwick Michael B Wilson Ian A |
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Institution: | 1 Department of Molecular Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA2 Department of Immunology and Microbial Science, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA3 Skaggs Institute for Chemical Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, CA 92037, USA |
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Abstract: | Antibody Z13e1 is a relatively broadly neutralizing anti-human immunodeficiency virus type 1 antibody that recognizes the membrane-proximal external region (MPER) of the human immunodeficiency virus type 1 envelope glycoprotein gp41. Based on the crystal structure of an MPER epitope peptide in complex with Z13e1 Fab, we identified an unrelated protein, interleukin (IL)-22, with a surface-exposed region that is structurally homologous in its backbone to the gp41 Z13e1 epitope. By grafting the gp41 Z13e1 epitope sequence onto the structurally homologous region in IL-22, we engineered a novel protein (Z13-IL22-2) that contains the MPER epitope sequence for use as a potential immunogen and as a reagent for the detection of Z13e1-like antibodies. The Z13-IL22-2 protein binds Fab Z13e1 with a Kd of 73 nM. The crystal structure of Z13-IL22-2 in complex with Fab Z13e1 shows that the epitope region is faithfully replicated in the Fab-bound scaffold protein; however, isothermal calorimetry studies indicate that Fab binding to Z13-IL22-2 is not a lock-and-key event, leaving open the question of whether conformational changes upon binding occur in the Fab, in Z13-IL-22, or in both. |
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Keywords: | HIV-1 human immunodeficiency virus type 1 Env envelope CDR complementarity-determining region MPER membrane-proximal external region WT wild type IL interleukin mAb monoclonal antibody bNAb broadly neutralizing antibody PDB Protein Data Bank ITC isothermal titration calorimetry o/n overnight RT room temperature PBS phosphate-buffered saline TPBS PBS containing 0 05% Tween-20 NFDM nonfat dry milk |
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