Bradykinin B2 receptor-mediated transport into intact cells : anti-receptor antibody-based cargoes

Authors: Bawolak, Marie-ThérèseLodge, RobertMorissette, GuillaumeMarceau, François
Abstract: Endocytosis of the bradykinin-stimulated B2 receptors is parallel to the transport and subsequent degradation of the ligand. To implement biotechnological applications based on receptor-mediated transport, one strategy is to conjugate the agonist ligand to a cargo. Alternatively, we studied whether the B2 receptor can transport large antibody-based cargoes into intact cells and characterized the ensuing endosomal routing. Myc-tagged B2 receptors (coded by the vector myc-B2R) and a truncated construction devoid of the Ser–Thr phosphorylation domain (myc-B2Rtrunc vector) were coupled to anti-myc monoclonal antibodies that did not impair bradykinin binding or elicit calcium signaling in intact cells. Anti-myc antibodies, conjugated or not with secondary antibodies optionally coupled to Qdot nanomaterials, were transported into early endosome autoantigen 1-, and β-arrestin-positive vesicles in bradykinin-stimulated intact cells expressing receptors encoded by myc-B2R. Antibody-conjugated cargoes progressed into late-endosomes-lysosomes within 3 h without evidence of autophagy. Receptors encoded by myc-B2Rtrunc did not support the ligand-controlled endocytosis of anti-myc antibodies. Aside from small ligand-conjugated cargoes, very large antibody-based cargoes can be transported by agonist-stimulated B2 receptors into intact cells. The latter type of cargo requires a receptor competent for interaction with β-arrestins, enters the degradation pathway separately from the receptor as a function of time and has the potential to confer a qualitatively novel function to a receptor.
Document Type: Article de recherche
Issue Date: 7 July 2011
Open Access Date: 21 November 2017
Document version: AM
This document was published in: European Journal of Pharmacology, Vol. 668 (1-2), 107–114 (2011)
Alternative version: 10.1016/j.ejphar.2011.06.041
Collection:Articles publiés dans des revues avec comité de lecture

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