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• W2060319767 endingPage "e44669" @default.
• W2060319767 startingPage "e44669" @default.
• W2060319767 abstract "Stimulating, and some blocking, antibodies to the TSH receptor (TSHR) have conformation-dependent epitopes reported to involve primarily the leucine rich repeat region of the ectodomain (LRD). However, successful crystallization of TSHR residues 22–260 has omitted important extracellular non-LRD residues including the hinge region which connects the TSHR ectodomain to the transmembrane domain and which is involved in ligand induced signal transduction. The aim of the present study, therefore, was to determine if TSHR antibodies (TSHR-Abs) have non-LRD binding sites outside the LRD. To obtain this information we employed the method of epitope protection in which we first protected TSHR residues 1–412 with intact TSHR antibodies and then enzymatically digested the unprotected residues. Those peptides remaining were subsequently delineated by mass spectrometry. Fourteen out of 23 of the reported stimulating monoclonal TSHR-Ab crystal contact residues were protected by this technique which may reflect the higher binding energies of certain residues detected in this approach. Comparing the protected epitopes of two stimulating TSHR-Abs we found both similarities and differences but both antibodies also contacted the hinge region and the amino terminus of the TSHR following the signal peptide and encompassing cysteine box 1 which has previously been shown to be important for TSH binding and activation. A monoclonal blocking TSHR antibody revealed a similar pattern of binding regions but the residues that it contacted on the LRD were again distinct. These data demonstrated that conformationally dependent TSHR-Abs had epitopes not confined to the LRDs but also incorporated epitopes not revealed in the available crystal structure. Furthermore, the data also indicated that in addition to overlapping contact regions within the LRD, there are unique epitope patterns for each of the antibodies which may contribute to their functional heterogeneity." @default.
• W2060319767 created "2016-06-24" @default.
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• W2060319767 date "2012-09-05" @default.
• W2060319767 modified "2023-09-26" @default.
• W2060319767 title "Antibody Protection Reveals Extended Epitopes on the Human TSH Receptor" @default.
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• W2060319767 doi "https://doi.org/10.1371/journal.pone.0044669" @default.
• W2060319767 hasPubMedCentralId "https://www.ncbi.nlm.nih.gov/pmc/articles/3434159" @default.
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• W2060319767 hasPublicationYear "2012" @default.
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