Characterization of Putative CLC2 Interacting Proteins

• Main Authors: Cheng-Hsin Liu, 劉政昕
• Other Authors: Chih-Yung Tang
• Format: Others
• Language: zh-TW
• Published: 2011
• Online Access: http://ndltd.ncl.edu.tw/handle/70526399850207149236

碩士 === 國立臺灣大學 === 生理學研究所 === 99 === CLC2 channel is a ubiquitously expressed voltage-gated chloride channel that is activated by hyperpolarization, cell swelling, extracellular acidification, and rise of intracellular chloride concentration. In recent studies, CLC2 has been suggested to assist chloride extrusion in hippocampal pyramidal cells. In addition, CLC2 has been implicated to have neuroprotective roles in aging mice. However, the precise cellular regulatory properties of CLC2 channel in neurons remain elusive. Therefore, we aimed to understand the physiological roles and cellular signaling mechanisms of CLC2 channels by searching for novel CLC2-interacting proteins. Previous studies in our lab utilizing yeast-two hybrid screening of rat brain cDNA library has identified several potential CLC2-interacting proteins. In this study, we further confirmed these interactions by using co-immunoprecipitation and GST pull-down assay. Our data suggest that four of these candidate proteins display significant interaction with CLC2. Immunofluorescence analysis revealed that these candidate proteins failed to affect the subcellular localization of CLC2 in HEK293T cells. It is still unclear whether total CLC2 expression may be affected in the presence of candidate proteins and thus required further studies. In the future, we plan to apply electrophysiological and biochemical assays to determine whether these interacting candidates can affect the biophysical and biosynthetic properties of CLC2 channels.