Regulation of the Water Channel Aquaporin-2 via 14-3-3θ and -ζ

Moeller, Hanne B.; Slengerik-Hansen, Joachim; Aroankins, Takwa S.; Assentoft, Mette; MacAulay, Nanna; Moestrup, Søren K.; Bhalla, Vivek; Fenton, Robert A.

doi:10.1074/jbc.m115.691121

Cited by 33 publications

(32 citation statements)

References 50 publications

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“…44 mpkCCD14 Culture and Experimental Conditions for CHIP Analyses mpkCCD14 cells were cultured on semipermeable supports as previously described. 45 For acute dDAVP stimulation, cells were washed twice in pure media and reincubated in pure media for 3 hours before being restimulated with dDAVP (1 nM) from the basolateral side for 20 minutes at 37°C. In dDAVP washout experiments, cells were subsequently washed twice in pure media and reincubated for 30 minutes at 37°C before sample preparation.…”

Section: Real-time Quantitative Pcrmentioning

confidence: 99%

CHIP Regulates Aquaporin-2 Quality Control and Body Water Homeostasis

Moeller

Stevens

et al. 2017

JASN

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The importance of the kidney distal convoluted tubule (DCT) and cortical collecting duct (CCD) is highlighted by various water and electrolyte disorders that arise when the unique transport properties of these segments are disturbed. Despite this critical role, little is known about which proteins have a regulatory role in these cells and how these cells can be regulated by individual physiologic stimuli. By combining proteomics, bioinformatics, and cell biology approaches, we found that the E3 ubiquitin ligase CHIP is highly expressed throughout the collecting duct; is modulated in abundance by vasopressin; interacts with aquaporin-2 (AQP2), Hsp70, and Hsc70; and can directly ubiquitylate the water channel AQP2 shRNA knockdown of CHIP in CCD cells increased AQP2 protein and reduced AQP2 ubiquitylation, resulting in greater levels of AQP2 and phosphorylated AQP2. CHIP knockdown increased the plasma membrane abundance of AQP2 in these cells. Compared with wild-type controls, CHIP knockout mice or novel CRISPR/Cas9 mice without CHIP E3 ligase activity had greater AQP2 abundance and altered renal water handling, with decreased water intake and urine volume, alongside higher urine osmolality. We did not observe significant changes in other water- or sodium-transporting proteins in the gene-modified mice. In summary, these results suggest that CHIP regulates AQP2 and subsequently, renal water handling.

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Section: Real-time Quantitative Pcrmentioning

confidence: 99%

CHIP Regulates Aquaporin-2 Quality Control and Body Water Homeostasis

Moeller

Stevens

et al. 2017

JASN

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“…The 14 -3-3 proteins, which are phospho-serine/phosphothreonine binding proteins (1,85), have also been reported as AQP2-binding partners, depending on the phosphorylation of AQP2 (84). The levels of two isoforms of 14 -3-3 proteins, 14 -3-3 and 14 -3-3 were increased by short-term vasopressin stimulation.…”

Section: Aqp2-binding Protein Complex In Aqp2 Traffickingmentioning

confidence: 99%

“…The levels of two isoforms of 14 -3-3 proteins, 14 -3-3 and 14 -3-3 were increased by short-term vasopressin stimulation. However, the levels of ubiquitination, half-life, and the expression levels of AQP2 were changed differently, when each isoform was knock-downed, separately (84). Moreover, in the degradation process of AQP2, lysosomal trafficking regulator-interacting protein 5 (LIP5) interacted with the carboxy-terminal region of AQP2, independent on phosphorylation (S256) and ubiquitination (K270) of AQP2 (146).…”

Section: Aqp2-binding Protein Complex In Aqp2 Traffickingmentioning

confidence: 99%

Molecular mechanisms regulating aquaporin-2 in kidney collecting duct

Jung

Kwon

2016

American Journal of Physiology-Renal Physiology

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The kidney collecting duct is an important renal tubular segment for regulation of body water homeostasis and urine concentration. Water reabsorption in the collecting duct principal cells is controlled by vasopressin, a peptide hormone that induces the osmotic water transport across the collecting duct epithelia through regulation of water channel proteins aquaporin-2 (AQP2) and aquaporin-3 (AQP3). In particular, vasopressin induces both intracellular translocation of AQP2-bearing vesicles to the apical plasma membrane and transcription of the Aqp2 gene to increase AQP2 protein abundance. The signaling pathways, including AQP2 phosphorylation, RhoA phosphorylation, intracellular calcium mobilization, and actin depolymerization, play a key role in the translocation of AQP2. This review summarizes recent data demonstrating the regulation of AQP2 as the underlying molecular mechanism for the homeostasis of water balance in the body.

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“…11 Direct interaction between AQPs and a regulatory protein has mainly been probed using AQP peptides corresponding to putative binding sites, 5,6,12 which oen gives valuable information concerning location of binding sites and specic interacting residues but does not provide the complete biological context. Peptide-based interaction studies may therefore miss additional factors that affect binding affinity, such as allosteric sites and/or the accessibility of the interaction motif.…”

Section: Introductionmentioning

confidence: 99%

Protein–protein interactions in AQP regulation – biophysical characterization of AQP0–CaM and AQP2–LIP5 complex formation

et al. 2018

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Protein-protein interactions play important roles in regulating human aquaporins (AQP) by gating as well as trafficking. While structural and functional studies have provided detailed knowledge of AQP transport mechanisms, selectivity as well as gating by conformational changes of loops or termini, the mechanism behind how protein-protein interactions control AQP-mediated water transport through cellular membranes remains poorly characterized. Here we explore the interaction between two human AQPs and regulatory proteins: the interaction between AQP0 and calmodulin, which mediates AQP0 gating, as well as the interaction between AQP2 and LIP5, which is involved in trafficking. Using microscale thermophoresis (MST) and fluorescence anisotropy, two methods that have the advantage of low sample consumption and detergent compatibility, we show that the interactions can be studied using both full-length AQPs and AQP peptides corresponding to the regulatory protein binding sites. However, full-length AQPs gave better reproducibility between methods and for the first time revealed that AQP0 binds CaM in a cooperative manner, which was not seen in experiments using peptides. Our study highlights that, while peptides are great tools for locating binding sites and pinpointing interacting residues, fulllength proteins may give additional insights, such as binding mechanism, allostery and cooperativity, important parameters for understanding protein-protein mediated regulation in the cellular context. Our work provides a platform for further studies of AQP regulation that may be of interest for designing drugs that target AQP complexes as well as the development of artificial bio-mimetic water channels for water-purification purposes.

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Regulation of the Water Channel Aquaporin-2 via 14-3-3θ and -ζ

Cited by 33 publications

References 50 publications

CHIP Regulates Aquaporin-2 Quality Control and Body Water Homeostasis

CHIP Regulates Aquaporin-2 Quality Control and Body Water Homeostasis

Molecular mechanisms regulating aquaporin-2 in kidney collecting duct

Protein–protein interactions in AQP regulation – biophysical characterization of AQP0–CaM and AQP2–LIP5 complex formation

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