2026-03-17T00:28:13Zhttps://uvadoc.uva.es/oai/request

oai:uvadoc.uva.es:10324/475012022-07-18T09:33:04Zcom_10324_32522com_10324_952com_10324_894col_10324_32523

Chao, Ta-Hsiang Ember, Julia A. Wang, Meiying Bayón Prieto, Yolanda Hugli, Tony E. Ye, Richard D. 2021-07-19T06:09:55Z 2021-07-19T06:09:55Z 1999 Journal of Biological Chemistry, 1999, vol. 274, n. 14, p. 9721-9728 0021-9258 https://uvadoc.uva.es/handle/10324/47501 10.1074/jbc.274.14.9721 9721 14 9728 Journal of Biological Chemistry 274 The C3a anaphylatoxin receptor (C3aR) is a G protein-coupled receptor with an unusually large second extra-cellular loop (e2 loop,;172 amino acids). To determinethe function of this unique structure, chimeric and de-letion mutants were prepared and analyzed in trans-fected RBL-2H3 cells. Whereas replacement of the C3aRN-terminal segment with that from the human C5a re-ceptor had minimal effect on C3a binding, substitutionof the e2 loop with a smaller e2 loop from the C5a recep-tor (C5aR) abolished binding of125I-C3a and C3a-stimu-lated calcium mobilization. However, as much as 65% ofthe e2 loop sequence (amino acids 198 –308) may be re-moved without affecting C3a binding or calcium re-sponses. The e2 loop sequences adjacent to the trans-membrane domains contain multiple aspartate residuesand are found to play an important role in C3a bindingbased on deletion mutagenesis. Replacement of five as-partate residues in the e2 loop with lysyl residues sig-nificantly compromised both the binding and functionalcapabilities of the C3a receptor mediated by intact C3aor by two C3a analog peptides. These data suggest atwo-site C3a-C3aR interaction model similar to that es-tablished for C5a/C5aR. The anionic residues near the Nand C termini of the C3aR e2 loop constitute a non-effector secondary interaction site with cationic resi-dues in the C-terminal helical region of C3a, whereas theC3a C-terminal sequence LGLAR engages the primaryeffector site in C3aR. eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-nd/4.0/ © Elsevier Attribution-NonCommercial-NoDerivatives 4.0 Internacional Role of the second extracellular loop of human C3a receptor in agonist binding and receptor function info:eu-repo/semantics/article