Molecular Cloning of the Receptor for Human Antidiuretic Hormone

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Title: Molecular Cloning of the Receptor for Human Antidiuretic Hormone
Authors: Birnbaumer, Mariel; Seibold, Anita; Gilbert, Stephanie; Ishido, Ph.D., Masami; Barberis, Claude; Antaramian, Anaid; Brabet, Philippe; Rosenthal, Walter
Publisher: Nature
Date Published: May 28, 1992
Reference Number: 432
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Antidiuresis, the recovery of water from the lumen of the renal collecting tubule, is regulated by the hypothalamic release of antidiuretic hormone (ADH), which binds to specific receptors on renal collecting tubule cells, stimulates adenylyl cyclase and promotes the cyclic AMP-mediated incorporation of water pores into the luminal surface of these cells. We report here the isolation of the human ADH receptor gene using a genomic expression cloning approach. The gene was used to clone the complementary DNA from a human renal library. The deduced amino-acid sequence of the receptor yields a hydropathy profile characteristic of receptors with seven putative transmembrane regions. This and the comparison with other cloned receptors indicates that the ADH receptor is a member of the superfamily of G-protein-coupled receptors.

This translation by the NDI Foundation is to assist the lay reader. To provide a clear, accessible interpretation of the original article, we eliminated or simplified some technical detail and complicated scientific language. We concentrated our translation on those aspects of the article dealing directly with NDI. The NDI Foundation thanks the researchers for their work toward understanding and more effectively treating this disorder.
© Copyright NDI Foundation 2007 (JC)

A receptor is a molecular structure that binds with an extracellular messenger such as a hormone. Once the hormone binds with its receptor, its hormonal message is passed on through a molecular sequence setting in motion a specific physiologic response. Receptors are specific; they bind only with certain messengers. For example, a vasopressin receptor will only bind with the vasopressin hormone.

Birnbaumer, et al., report on their successful cloning of the gene which synthesizes one specific receptor that the antidiuretic hormone, arginine vasopressin (AVP) binds with. By binding with this specific receptor, which is located in the principal cells of the kidney collecting duct, AVP allows the kidneys to concentrate urine and reabsorb water flowing through the kidney collecting duct. The molecular sequence that allows the kidneys to do this is as follows: AVP binds with its receptor. This stimulates the enzyme, adenylyl cyclase. This elevates cAMP, and cAMP helps direct water-transporting proteins to insert themselves in the apex of the membranes of the collecting duct principal cells. This makes the membranes more water permeable. And this is what allows urine concentration and water reabsorption.

To clone the receptor gene, the authors first constructed a set of cloned DNA fragments. They screened the clones for their ability to be responsive to AVP. This was measured by the amount of AVP binding sites per cell. They also measured the clones that created cell responsiveness to AVP for their ability to affect cells in such a way as to stimulate adenylyl cyclase. Both these screenings indicate the presence in the clones of the AVP receptor.

Testing the degrees to which AVP and its analogs stimulated the cells containing the AVP receptor indicated that the AVP receptor was the vasopressin-2 receptor (V2R). The V2R is predicted to be 371 amino acids long with a relative molecular mass of 40,285. It is a member of the superfamily of G-protein-coupled receptors.