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Signaling properties of human α1D-adrenoceptors lacking the carboxyl terminus: intrinsic activity, agonist-mediated activation, and desensitization

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Abstract

α1-Adrenoceptors are differentially regulated by protein kinase C-mediated phosphorylation. The most sensitive member of this family is the α1D-subtype, which is also characterized by a constitutive activity and a reduced expression at the plasma membrane controlled by the amino terminus. Information on the structural domains that determine the function and regulation of this receptor subtype is scarce. Therefore, the function and phosphorylation of amino terminus-truncated (Δ1–79, (ΔN)) α1D-adrenoceptors were studied and compared with those of α1D-adrenoceptors truncated both at the amino and carboxyl termini (Δ1–79 and Δ441–572, (ΔN–ΔC)). These receptors were stably expressed in rat-1 fibroblast, at relatively high density (≈2 pmol/mg of membrane protein), and showed intrinsic activity that was markedly increased by noradrenaline. Interestingly, activation of protein kinase C markedly attenuated (desensitized) the function of both ΔN and ΔN–ΔC α1D-adrenoceptors. These receptors were photolabeled and immunoprecitated with an antibody directed against an influenza hemagglutinin epitope inserted at the amino termini. Metabolic labeling with radioactive phosphate and receptor immunoprecipitation studies indicated that these receptors are phosphoproteins whose phosphorylation state is increased by noradrenaline and by activation of protein kinase C. Our data indicate that carboxyl terminus-truncated α1D-adrenoceptors are fully functional and subjected to regulation by phosphorylation. The roles of the carboxyl termini differ among α1-adrenoceptor subtypes.

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Acknowledgments

This work was supported by grants from the Consejo Nacional de Ciencia y Tecnología (79908) and Dirección General de Asuntos del Personal Académico (IN212609). The authors want to express their gratitude to Dr. José Vázquez-Prado for reading and correcting the manuscript and for suggesting valuable experiments.

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Authors and Affiliations

  1. Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Apartado Postal 70-248, Mexico, 04510, Mexico

    C. Ekaterina Rodríguez-Pérez, M. Teresa Romero-Ávila & J. Adolfo García-Sáinz

  2. Departamento de Biología Celular, Centro de Investigación y de Estudios Avanzados—Instituto Politécnico Nacional, CINVESTAV-IPN, Apdo. Postal 14-740, 07000, Mexico, Mexico

    Guadalupe Reyes-Cruz

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  1. C. Ekaterina Rodríguez-Pérez
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  2. M. Teresa Romero-Ávila
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  3. Guadalupe Reyes-Cruz
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  4. J. Adolfo García-Sáinz
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Correspondence to J. Adolfo García-Sáinz.

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Rodríguez-Pérez, C.E., Romero-Ávila, M.T., Reyes-Cruz, G. et al. Signaling properties of human α1D-adrenoceptors lacking the carboxyl terminus: intrinsic activity, agonist-mediated activation, and desensitization. Naunyn-Schmied Arch Pharmacol 380, 99–107 (2009). https://doi.org/10.1007/s00210-009-0428-6

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  • DOI: https://doi.org/10.1007/s00210-009-0428-6

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