Abstract
Vasoactive intestinal peptide (VIP) is a widespread neuropeptide, which exerts many biological functions through interaction with the VPAC1 receptor, a class II G protein-coupled receptor. Photoaffinity labeling studies combined with 3D molecular modeling demonstrated that the central and C-terminal parts of VIP (segment 6–28) have physical contacts with the N-terminal ectodomain (N-Ted) of VPAC1 receptor. However, the domain of the hVPAC1 receptor interacting with the N-terminus of VIP (1–5) is still unknown. We have synthesized a photoreactive probe Bpa0-VIP. After photolabeling and receptor cleavage, Nu-PAGE analysis revealed a 5-kDa labeled fragment corresponding to the 130–137 sequence of hVPAC1 receptor, indicating that the N-terminus of VIP also interacts with the N-ted. A photoreactive probe, Bpa0-PG97–269, was also synthesized with the specific peptide antagonist PG97–269. After photoaffinity labeling, a glycosylated 15-kDa fragment is identified by cyanogen bromide (CNBr) cleavage and corresponds to the 43–66 sequence of the hVPAC1 receptor N-ted. These results indicate that: (1) the N-terminal part of VIP physically interacts with the N-ted in the continuity of 6–28 VIP sequence; (2) the N-terminal part of VIP and the selective peptide antagonist (PG97–269) have different sites of interaction with the VPAC1 receptor N-ted.
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Acknowledgements
This research was supported by the Institut National de la Santé et de la Recherche Médicale, the Université Paris-Diderot (Paris 7) and also by a grant from Association de Recherche sur la Polyarthrite. E. Ceraudo is supported by grant FDT20070910820 from the Fondation pour la Recherche Médicale and by an « Interuniversity Poles of Attraction Program » (PAI) from Université Libre de Bruxelles (ULB), Belgium.
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Ceraudo, E., Tan, YV., Nicole, P. et al. The N-Terminal Parts of VIP and Antagonist PG97–269 Physically Interact with Different Regions of the Human VPAC1 Receptor. J Mol Neurosci 36, 245–248 (2008). https://doi.org/10.1007/s12031-008-9073-7
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DOI: https://doi.org/10.1007/s12031-008-9073-7