Mol Cell Biol. 2006 Jan;26(1):303-12.
Direct interaction with Rab11a targets the epithelial Ca2+ channels TRPV5 and TRPV6 to the plasma membrane.
van de Graaf, S. F., Chang, Q., Mensenkamp, A. R., Hoenderop, J. G., Bindels, R. J.,
["Department of Physiology, Nijmegen Centre for Molecular Life Sciences, Radboud University, Nijmegen, The Netherlands."]
["Department of Physiology, Nijmegen Centre for Molecular Life Sciences, Radboud University, Nijmegen, The Netherlands."]
TRPV5 and TRPV6 are the most Ca2+-selective members of the transient receptor potential (TRP) family of cation channels and play a pivotal role in the maintenance of Ca2+ balance in the body. However, little is known about the mechanisms controlling the plasma membrane abundance of these channels to regulate epithelial Ca2+ transport. In this study, we demonstrated the direct and specific interaction of GDP-bound Rab11a with TRPV5 and TRPV6. Rab11a colocalized with TRPV5 and TRPV6 in vesicular structures underlying the apical plasma membrane of Ca2+-transporting epithelial cells. This GTPase recognized a conserved stretch in the carboxyl terminus of TRPV5 that is essential for channel trafficking. Furthermore, coexpression of GDP-locked Rab11a with TRPV5 or TRPV6 resulted in significantly decreased Ca2+ uptake, caused by diminished channel cell surface expression. Together, our data demonstrated the important role of Rab11a in the trafficking of TRPV5 and TRPV6. Rab11a exerts this function in a novel fashion, since it operates via direct cargo interaction while in the GDP-bound configuration.
PMID: 16354700

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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 | |||
TRPV5 |
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Rab11a | Inference | Prediction | 16354700 | |||||
TRPV6 |
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Rab11a | Yeast two-hybrid | Mouse | C-terminus | Mouse | Kidney | cDNA library | 16354700 |
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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Validation: In vitro validation
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Assay with recombinant proteins | Reference | |||||||||
TRP channel construct | Interactor construct | |||||||||
TRP channel | Interactor | Method | Species | Region | Expression system | Species | Region | Expression system | ||
TRPV5 |
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Rab11a | Fusion protein-pull down assay | Mouse | Full-length | E. coli | Mouse | Full-length | In vitro translation | 16354700 |
TRPV6 |
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Rab11a | Fusion protein-pull down assay | Mouse | Full-length | E. coli | Mouse | Full-length | In vitro translation | 16354700 |
(
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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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 | ||||
TRPV5 |
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Rab11a | Co-immunoprecipitation | Xenopus oocyte | Mouse | Full-length | Mouse | Full-length | 16354700 | |
TRPV5 |
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Rab11a | Co-immunofluorescence staining | Xenopus oocyte | Mouse | Full-length | Mouse | Full-length | 16354700 | |
TRPV5 |
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Rab11a | Co-immunofluorescence staining | Mouse kidney section | 16354700 | |||||
TRPV6 |
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Rab11a | Co-immunofluorescence staining | Mouse kidney section | 16354700 | |||||
TRPV6 |
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Rab11a | Co-immunofluorescence staining | Xenopus oocyte | Mouse | Full-length | Mouse | Full-length | 16354700 |
(
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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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 | ||||
TRPV5 |
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Rab11a | Fusion protein-pull down assay | Mouse | 596-600 | Mouse | Not determined | 16354700 |
(
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click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)

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Functional consequence
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TRP channel | Interactor | Method | Post-translational modification | Subcellular trafficking | Activity | Reference | ||||||
TRPV5 |
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Rab11a | Co-immunofluorescence staining | Increase in plasma membrane level | 16354700 | |||||||
TRPV5 |
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Rab11a | Calcium measurement | Activation | 16354700 | |||||||
TRPV6 |
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Rab11a | Co-immunofluorescence staining | Increase in plasma membrane level | 16354700 | |||||||
TRPV6 |
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Rab11a | Calcium measurement | Activation | 16354700 |
(
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
