Specific binding activities and cyclic GMP responses by atrial natriuretic polypeptide in kidney epithelial cell line (LLC-PK1) |
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| Authors: | K Inui H Saito Y Matsukawa K Nakao N Morii H Imura M Shimokura Y Kiso R Hori |
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| Affiliation: | 1. Department of Pharmacy, Kyoto University Hospital, Sakyo-ku, Kyoto, 606, Japan;2. Second Division, Department of Medicine, Faculty of Medicine, Kyoto University, Sakyo-ku, Kyoto, 606, Japan;3. Radioisotope Research Center, Kyoto University, Sakyo-ku, Kyoto, 606, Japan;4. Department of Medicinal Chemistry, Kyoto Pharmaceutical University, Yamashina-ku, Kyoto, 607, Japan;1. Centre hospitalo-universitaire de Lille, hôpital C-Huriez, service d’endocrinologie et métabolisme, 1, rue Polonovski, 59037 Lille cedex, France;2. UMR 1190 recherche translationnelle sur le diabète Inserm, 59000 Lille, France;3. EGID (European Genomic Institute for Diabetes), université de Lille, 59000 Lille, France;1. Department of Cardiovascular Medicine, Kyoto University Graduate School of Medicine, Japan;2. Department of Internal Medicine, Wakakusa-Tatsuma Rehabilitation Hospital, Japan;3. Department of Cardiovascular Medicine, Shinshu University School of Medicine, Japan |
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| Abstract: | Receptor binding activities and cyclic GMP responses by alpha-human atrial natriuretic polypeptide (alpha-hANP) and its fragments were studied in a kidney epithelial cell line (LLC-PK1). Binding of 125I-alpha-hANP to the cells at 0 degrees C was saturable, time-dependent and reversible, indicating the presence of a single class of binding sites. alpha-hANP (7-23)NH2 fragment inhibited most effectively the specific binding of 125I-alpha-hANP to the LLC-PK1 cells, followed by alpha-hANP (17-28) and alpha-hANP (8-22), while alpha-hANP (1-6) and alpha-hANP (24-28) did not. alpha-hANP stimulated the formation of cyclic GMP in the LLC-PK1 cells dose-dependently. Although no fragments of alpha-hANP used were effective for cyclic GMP formation in the LLC-PK1 cells, alpha-hANP (7-23) NH2 antagonized the action of alpha-hANP on cyclic GMP formation. These data suggest that the LLC-PK1 cells retain specific receptors for atrial natriuretic polypeptide (ANP) and respond to ANP by stimulating cyclic GMP formation, and therefore this cell line may be useful for studying the mechanism of action for ANP in renal tubular cells. |
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