• Port-a-Patch

    世界上最小的膜片钳系统
  • Port-a-Patch

    易学易用——教学的理想工具
  • Port-a-Patch

    可记录细胞、细胞器与脂双层
  • Port-a-Patch

    世界上首款平板芯片膜片钳设备
  • Port-a-Patch

    可进行内灌流——更换细胞内液的理想工具

2015 - Different Ligands of the TRPV3 Cation Channel Cause Distinct Conformational Changes As Revealed by Intrinsic Tryptophan Fluorescence Quenching

icon pap  Port-a-Patch and   icon vpp   Vesicle Prep Pro publication in Journal of Biological Chemistry (2015)

Authors: 
Billen B., Brams M., Debaveye S., Remeeva A., Alpizar Y.A., Waelkens E., Kreir M., Brüggemann A., Talavera K., Nilius B., Voets T., Ulens C.

 

Journal: 
J Biol Chem. (2015) 290:12964-12974


Abstract: 

TRPV3 is a thermosensitive ion channel primarily expressed in epithelial tissues of the skin, nose, and tongue. The channel has been implicated in environmental thermosensation, hyperalgesia in inflamed tissues, skin sensitization, and hair growth. Although transient receptor potential (TRP) channel research has vastly increased our understanding of the physiological mechanisms of nociception and thermosensation, the molecular mechanics of these ion channels are still largely elusive. In order to better comprehend the functional properties and the mechanism of action in TRP channels, high-resolution three-dimensional structures are indispensable, because they will yield the necessary insights into architectural intimacies at the atomic level. However, structural studies of membrane protein c are currently hampered by difficulties in protein purification and in establishing suitable crystallization conditions. In this report, we present a novel protocol for the purification of membrane proteins, which takes advantage of a C-terminal GFP fusion. Using this protocol, we purified human TRPV3. We show that the purified protein is a fully functional ion channel with properties akin to the native channel using planar patch clamp on reconstituted channels and intrinsic tryptophan fluorescence spectroscopy. Using intrinsic tryptophan fluorescence spectroscopy, we reveal clear distinctions in the molecular interaction of different ligands with the channel. Altogether, this study provides powerful tools to broaden our understanding of ligand interaction with TRPV channels, and the availability of purified human TRPV3 opens up perspectives for further structural and functional studies.


Download here

返回总览

We use cookies on our website. Some of them are essential for the operation of the site, while others help us to improve this site and the user experience (tracking cookies). You can decide for yourself whether you want to allow cookies or not. Please note that if you reject them, you may not be able to use all the functionalities of the site.