Animal Cells Syst (Seoul). 2011 Jun;15(2):95-106. Epub 2011 Jun 9.
Mutation of a putative S-nitrosylation site of TRPV4 protein facilitates the channel activates.
Lee, E. J., Shin, S. H., Hyun, S., Chun, J., Kang, S. S.,
["Department of Pre-medicine, Eulji University School of Medicine, Daejeon 301-832, Republic of Korea."]
["Department of Pre-medicine, Eulji University School of Medicine, Daejeon 301-832, Republic of Korea."]
The transient receptor potential vanilloid 4 (TRPV4) cation channel, a member of the TRP vanilloid subfamily, is expressed in a broad range of tissues. Nitric oxide (NO) as a gaseous signal mediator shows a variety of important biological effects. In many instances, NO has been shown to exhibit its activities via a protein S-nitrosylation mechanism in order to regulate its protein functions. With functional assays via site-directed mutagenesis, we demonstrate herein that NO induces the S-nitrosylation of TRPV4 Ca(2+) channel on the Cys(853) residue, and the S-nitrosylation of Cys(853) reduced its channel sensitivity to 4-alpha phorbol 12,13-didecanoate and the interaction between TRPV4 and calmodulin. A patch clamp experiment and Ca(2+) image analysis show that the S-nitrosylation of Cys(853) modulates the TRPV4 channel as an inhibitor. Thus, our data suggest a novel regulatory mechanism of TRPV4 via NO-mediated S-nitrosylation on its Cys(853) residue.
PMID: 21837266

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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 | ||
TRPV4 |
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Calmodulin | Fusion protein-pull down assay | Human | 718-871 | E. coli | Not used | HEK293T lysates | 21837266 |
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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 | ||||
TRPV4 |
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Calmodulin | Fusion protein-pull down assay | Human | 718-871 | Human | Not determined | 21837266 |
(
:
click the arrow icon to show interactions only between the corresponding TRP channel and the interactor)
