2004 Global Researcher Conference Proceeding

April 09 - 11, 2004

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Conference: 2004 Global Researcher Conference
Title: The novel Aquaporin-2 maturing protein 1 interacts with AQP2, inhibits its forskolin-induced translocation to the apical membrane, and reduces its expression
Authors: Boone, Michelle; van Balkom, Bas; Hendriks, Giel; Kamsteeg, Erik-Jan; de Mattia, Fabrizio; van der voorde, Anne; van Herp, Francois; van der Sluijs, Peter; Deen, Peter M.T.
Institutions: Radboud University Nijmegen Medical Center, University of Nijmegen, Utrecht University School of Medicine, Utrecht University
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Boone Translocation of the Aquaporin-2 (AQP2) water channel to and from the apical membrane is regulated by the antidiuretic hormone vasopressin (AVP). In states of hypernatremia or hypovolemia, binding of AVP to its type 2 receptor increases cAMP concentration through the activation of adenylate cyclase. Increases in the intracellular cAMP concentration results in the activation of PKA, leading to the phosphorylation of three to four monomers of the AQP2 homotetramer and the translocation of AQP2 to the apical membrane. Regulation of protein routing and expression involves the recognition of motifs in these proteins by interacting protein complexes. Recently we identified a first and novel protein of 42 kDa, which interacted with the C-terminal tail of AQP2 and which we named Aquaporin-2 maturing protein 1 (AMP1). Immunohistochemistry on kidney sections revealed that AMP1 and AQP2 were co-expressed in the same cells. Yeast-2-hybrid analysis showed that AMP1 interacted with the tail of AQP2, AQP2-S256A and AQP2-S256D, but not with those of AQP3 or AQP4. To unravel its function, AMP1 was co-expressed in wt10 cells, which are Madin Darby Canine Kidney (MDCK) cells, in which AQP2 is translocated from intracellular vesicles to the plasma membrane upon treatment with forskolin. Co-expression with AMP1 interfered with the forskolin-induced translocation of AQP2, which was not observed for AQP4. Furthermore, AMP1 was shown to reduce the expression of AQP2 in several clones. The decrease in AQP2 expression and disturbed sorting of AQP2 to the apical membrane as found with AMP1 expression are as found in acquired NDI. As such, AMP1 might have important role in the regulation of AQP2 in these forms of NDI.

The journey of AQP2 from the cell interior to the cell membrane is initiated by the binding of the antidiuretic hormone, arginine vasopressin (AVP), with the vasopressin–2 receptor (V2R), and is orchestrated by a cascade of molecular interactions set into motion by this binding. An emerging area of knowledge reveals that part of what regulates proteins such as AQP2 as it travels to the cell membrane is the interaction of other protein complexes that recognize certain configurations of the traveling protein.

Boone, et al., identified a protein that interacts with the tail end of the AQP2. They named it Aquaporin-2 maturing protein (AMP1). The researchers discovered that AMP1 and AQP2 are expressed in the same cells. To discover the function of AMP1, the researchers placed both proteins in a cell culture. Normally, AQP2 will move to the cell membrane when the chemical, forskolin, is added to the culture. However, AMP1 interfered with AQP2’s movement. Further, AMP1 seems to limit the amount of AQP2s that are produced. AMP1 produces the same effects on AQP2 that are found in acquired forms of NDI. The scientists speculate that AMP1 may play an important role in the regulation of AQP2 in these forms of NDI.