The human chorionic gonadotropin-ß arginine68 to glutamic acid substitution fixes the conformation of the C-terminal Peptide
Charrel-Dennis, Marie, Terrazzini, Nadia, McBride, Jeffrey D., Martensen, Pia M., Justesen, Just, Berger, Peter, Lapthorn, Adrian J., Kelly, Charles, Roitt, Ivan, Delves, Peter J. and Lund, Torben (2005) The human chorionic gonadotropin-ß arginine68 to glutamic acid substitution fixes the conformation of the C-terminal Peptide. Molecular Endocrinology, 19 (7). pp. 1803-1811. ISSN 0888-8809
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Wild-type human chorionic gonadotropin (hCG) has been used as a contraceptive vaccine. However, extensive sequence homology with LH elicits production of cross-reactive antibodies. Substitution of arginine68 of the ß-subunit (hCGß) with glutamic acid (R68E) profoundly reduces the cross-reactivity while refocusing the immune response to the hCGß-specific C-terminal peptide (CTP). To investigate the molecular basis for this change in epitope usage, we immunized mice with a plasmid encoding a truncated hCGß-R68E chain lacking the CTP. The animals produced LH-cross-reactive antibodies, suggesting that the refocused immunogenicity of R68E is a consequence of epitope masking by a novel disposition of the CTP in the mutant rather than a structural change in the cross-reactive epitope region. This explanation was strongly supported by surface plasmon resonance analysis using a panel of anti-hCGß-specific and anti-hCGß/LH cross-reactive monoclonal antibodies (mAbs). Whereas the binding of the LH cross-reactive mAbs to hCGß-R68E was eliminated, mAbs reacting with hCGß-specific epitopes bound to hCGß and hCGß-R68E with identical affinities. In a separate series of experiments, we observed that LH cross-reactive epitopes were silent after immunization with a plasmid encoding a membrane form of hCGß-R68E, as previously observed with the soluble mutant protein itself. In contrast, the plasmid encoding the soluble secreted form of hCGß-R68E evoked LH cross-reactive antibodies, albeit of relatively low titer, suggesting that the handling and processing of the proteins produced by the two constructs differed.
|Research Areas:||Science & Technology > Biomedical Science|
|Deposited On:||02 Feb 2010 09:05|
|Last Modified:||22 Oct 2013 07:41|
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