Clc - Proteoliposomes with different lipid-to-protein ratios
We tested proteoliposomes with different densities of reconstituted Clc protein for their signal-to-noise ratios and compared the obtained signal amplitudes with a negative control: liposomes without reconstituted protein.
Background and Assay Description:
We have performed a single solution exchange workflow to activate Clc by 50 mM Chloride concentration jump at pH 5.2. Each trace shown was recorded from a different sample. We have used proteoliposomes with different lipid-to-protein ratios. The higher the protein density (lower LPR), the higher the signal-to-noise ratio during the SURFE2R assay. In addition, current signals reflecting transport reactions decay faster, when the protein density is high, since more transport accelerates the charging of the membrane due to Chloride transport. As a control we performed the same assay using protein-free liposomes (red trace), which did not show any significant signal.
Author: Nanion Technologies, in collaboration with Merritt Maduke, Stanford University School of Medicine
Target: EriC, ClcA, ClC-ecl
Family / Type: Chloride Carrier/Channel Family (ClCF, TCDB: 2.A.49)
Mode of transport: H+/2Cl- exchange
Organism: Escherichia coli
Sample: Proteoliposomes from purified protein, LPR 20 – 2000 and liposomes without protein
Expression: Recombinant overexpression in E.coli
Platform: SURFE²R N1