8RTW
Crystal Structure of the Anti-testosterone Fab in Complex with Testosterone
Summary for 8RTW
| Entry DOI | 10.2210/pdb8rtw/pdb |
| Descriptor | Anti-testosterone Fab 220 light chain, Anti-testosterone Fab 220 heavy chain, TESTOSTERONE, ... (4 entities in total) |
| Functional Keywords | fab, testosterone, immune system |
| Biological source | Mus musculus More |
| Total number of polymer chains | 4 |
| Total formula weight | 97816.91 |
| Authors | Eronen, V.,Parkkinen, T.,Hakulinen, N.,Rouvinen, J. (deposition date: 2024-01-29, release date: 2024-09-18, Last modification date: 2024-11-13) |
| Primary citation | Eronen, V.,Takkinen, K.,Torni, A.,Peng, K.,Janis, J.,Parkkinen, T.,Hakulinen, N.,Rouvinen, J. Structural insights into ternary immunocomplex formation and cross-reactivity: binding of an anti-immunocomplex FabB12 to Fab220-testosterone complex. Febs J., 291:4744-4756, 2024 Cited by PubMed Abstract: Anti-immunocomplex (Anti-IC) antibodies have been used in developing noncompetitive immunoassays for detecting small molecule analytics (haptens). These antibodies bind specifically to the primary antibody in complex with hapten. Although several anti-IC antibody-based immunoassays have been developed, structural studies of these systems are very limited. In this study, we determined the crystal structures of anti-testosterone Fab220 in complex with testosterone and the corresponding anti-IC antibody FabB12. The structure of the ternary complex of testosterone, Fab220, and FabB12 was predicted using LightDock and AlphaFold. The ternary complex has a large (~ 1100 Å) interface between antibodies. The A-ring of the testosterone bound by Fab220 also participates in the binding of the anti-IC antibody. The structural analysis was complemented by native mass spectrometry. The affinities for testosterone (TES) and three cross-reactive steroids [dihydrotestosterone (DHT), androstenedione (A4), and dehydroepiandrosterone sulfate (DHEA-S)] were measured, and ternary complex formation was studied. The results clearly show the ternary complex formation in the solution. Although DHT showed significant cross-reactivity, A4 and DHEA-S exhibited minor cross-reactivity. PubMed: 39206623DOI: 10.1111/febs.17258 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.99 Å) |
Structure validation
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