J Biol Chem. 2004 Aug 13;279(33):35009-16. Epub 2004 Jun 12.
PIGEA-14, a novel coiled-coil protein affecting the intracellular distribution of polycystin-2.
Hidaka, S., Konecke, V., Osten, L., Witzgall, R.,
["Institute for Molecular and Cellular Anatomy, University of Regensburg, Universitatsstrasse 31, 93053 Regensburg, Germany."]
["Institute for Molecular and Cellular Anatomy, University of Regensburg, Universitatsstrasse 31, 93053 Regensburg, Germany."]
Employing a yeast two-hybrid screen with the COOH terminus of polycystin-2, one of the proteins mutated in patients with polycystic kidney disease, we were able to isolate a novel protein that we call PIGEA-14 (polycystin-2 interactor, Golgi- and endoplasmic reticulum-associated protein with a molecular mass of 14 kDa). Molecular modeling only predicts a coiled-coil motif, but no other functional domains, in PIGEA-14. In a subsequent two-hybrid screen using PIGEA-14 as a bait, we found GM130, a component of the cis-compartment of the Golgi apparatus. Co-expression of the PIGEA-14 and PKD2 cDNAs in LLC-PK(1) and HeLa cells resulted in a redistribution of PIGEA-14 and polycystin-2 to the trans-Golgi network, which suggests that PIGEA-14 plays an important role in regulating the intracellular location of polycystin-2 and possibly other intracellular proteins. Our results also indicate that the intracellular trafficking of polycystin-2 is regulated both at the level of the endo-plasmic reticulum and that of the trans-Golgi network.
PMID: 15194699
 Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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Screening
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| Experimental screening | Non-experimental screening | Reference | ||||||||
| TRP channel construct | Interactor source | |||||||||
| TRP channel | Interactor | Method | Species | Region | Species | Organ/tissue | Sample type | |||
| TRPP1 |
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PIGEA-14/CBY1 | Yeast two-hybrid | Human | 680-968 | Human | Kidney | cDNA library | 15194699 | |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
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Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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top
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| Validation: In vivo validation
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| Assay with endogenous proteins | Assay with overexpressed proteins | Reference | ||||||||
| Cell or tissue | Cell or tissue | TRP channel construct | Interactor construct | |||||||
| TRP channel | Interactor | Method | Species | Region | Species | Region | ||||
| TRPP1 |
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PIGEA-14/CBY1 | Co-immunoprecipitation | LLC-PK1 | Human | Full-length | Human | Full-length | 15194699 | |
| TRPP1 |
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PIGEA-14/CBY1 | Co-immunofluorescence staining | HeLa | Human | Full-length | Human | Full-length | 15194699 | |
(:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
|
Screening
Validation: In vitro validation
Validation: In vivo validation
Characterization
Functional consequence
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top
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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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PIGEA-14/CBY1 | Yeast two-hybrid | Human | 680-968 | Human | 60-112 | 15194699 | ||
| TRPP1 |
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PIGEA-14/CBY1 | Yeast two-hybrid | Human | 833-923 | Human | Not determined | 15194699 | ||
(:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)