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Identification of a Potential Receptor for Both Peptide Histidine Isoleucine and Peptide Histidine Valine

Department of Zoology (D.L.Y.T., R.T.K.P., A.O.L.W., S.M.C., B.K.C.C.), University of Hong Kong, Hong Kong, People’s Republic of China; and European Institute for Peptide Research (H.V.), INSERM Unité 413, UA Centre National de la Recherche Scientifique, University of Rouen,76821 Mont-Saint-Aignan, France

Address all correspondence and requests for reprints to: Dr. Billy K. C. Chow, Department of Zoology, The University of Hong Kong, Pokfulam Road, Hong Kong, SAR, PRC. E-mail: . bkcc{at}hkusua.hku.hk

Peptide histidine isoleucine (PHI), peptide histidine valine (PHV), and vasoactive intestinal polypeptide (VIP) are cosynthesized from the same precursor and share high levels of structural similarities with overlapping biological functions. In this study, the first PHI/PHV receptor was isolated and characterized in goldfish. To study this receptor using homologous peptides, we have also characterized the goldfish prepro-PHI/VIP, and, surprisingly, a shorter transcript lacking the VIP coding region was isolated. A PHI/VIP precursor without the VIP coding sequence has never before been reported. Initial functional expression of the PHI/PHV receptor in Chinese hamster ovary cells revealed that it could be activated by human PHV [50% effective concentration (EC₅₀): 43 nM] and to a lesser extent human PHI (EC₅₀: 133 nM) and helodermin (EC₅₀: 166 nM) but not fish and mammalian pituitary adenylate cyclase-activating polypeptides and VIPs. Subsequent studies indicated that, similar to the pituitary adenylate cyclase-activating polypeptide receptors (PAC1-R, VPAC1-R, and VPAC2-R), the receptor isolated in this study is able to interact with goldfish PHI and its C-terminally extended form, PHV with EC₅₀ values 93 and 43 nM, respectively. Northern blot and RT-PCR/Southern blot analyses revealed that the PHI/VIP gene is expressed in the intestine, brain, and gall bladder and the PHI/PHV receptor gene is primarily expressed in the pituitary and to a lesser extend in the intestine and gall bladder, suggesting that PHI/PHV may play a role, notably in the regulation of pituitary function. In conclusion, our results demonstrate for the first time the existence of a PHI/PHV receptor, indicating that the functions of PHI and PHV could be mediated by their own receptor in addition to VIP receptors.

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