Am J Physiol Renal Physiol. 2009 Nov;297(5):F1310-5. doi: 10.1152/ajprenal.00412.2009. Epub 2009 Sep 2.
Analysis of the cytoplasmic interaction between polycystin-1 and polycystin-2.
Casuscelli, J., Schmidt, S., DeGray, B., Petri, E. T., Celic, A., Folta-Stogniew, E., Ehrlich, B. E., Boggon, T. J.,
["Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520-8066, USA."]
["Department of Pharmacology, Yale University School of Medicine, New Haven, CT 06520-8066, USA."]
Autosomal dominant polycystic kidney disease (ADPKD) arises following mutations of either Pkd1 or Pkd2. The proteins these genes encode, polycystin-1 (PC1) and polycystin-2 (PC2), form a signaling complex using direct intermolecular interactions. Two distinct domains in the C-terminal tail of PC2 have recently been identified, an EF-hand and a coiled-coil domain. Here, we show that the PC2 coiled-coil domain interacts with the C-terminal tail of PC1, but that the PC2 EF-hand domain does not. We measured the K0.5 of the interaction between the C-terminal tails of PC1 and PC2 and showed that the direct interaction of these proteins is abrogated by a PC1 point mutation that was identified in ADPKD patients. Finally, we showed that overexpression of the PC1 C-terminal tail in MDCK cells alters the Ca2+ response, but that overexpression of the PC1 C-terminal tail containing the disease mutation does not. These results allow a more detailed understanding of the mechanism of pathogenic mutations in the cytoplasmic regions of PC1 and PC2.
PMID: 19726544
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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Characterization
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| Binding region mapping | Stoichiometry | Affinity (Kd) | Reference | |||||||
| TRP channel | Interactor | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPP1 |
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PKD1/Polycystin-1 | Fusion protein-pull down assay | Human | 821-927 | Mouse | 4183-4270 | 19726544 | ||
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