CIC - "Electrophysiological recordings of the H+/ Cl- exchanger Ec-ClC on the SURFE2R N1"
SURFE2R N1 application note: (0.3 MB)
Samples kindly provided by Dr. Merritt Maduke, Stanford University, USA
ClCs are a family of chloride ion channels and transporters with important physiological roles including regulation of the membrane potential, transepithelial salt transport and ion homeostasis1,2. To date, 9 members of the ClC family have been identified in mammals1,2, the first 4 (ClC-1, ClC-2, ClC-Ka, and ClC-Kb) are located on the plasma membrane where they act as chloride ion channels whereas the remaining 5 are located in intracellular organelles (ClC-3-7) and are chloride-proton exchangers1-3. These transporters are important for endosome, lysosome and synaptic vesicle acidification1,2, and mutations in, e.g. ClC-5 underlie the rare chronic kidney disorder, Dent’s Disease, and mutations in ClC-7 underlie osteopetrosis 2,4, a rare inherited bone hardening disorder. Given their ubiquitous expression and importance in physiological processes, they are important potential drug targets.
ClC from Escherichia coli (Ec-ClC or ClC-ec1) is closely ClC from Escherichia coli (Ec-ClC or ClC-ec1) is closely related to its mammalian counterparts and is a Cl-/H+ exchanger3,5. It transports 2 Cl- into the cell, coupled to the efflux of 1 H+3,5. Therefore Ec-ClC is an electrogenic transporter, generating a net charge flow. In E. coli, the Ec-ClC mediates acid resistance of enteric bacteria by promoting H+ extrusion1,5,6. Here we present Ec-ClC activity measurements on the SURFE2R N1 instrument using proteoliposomes reconstituted with purified Ec-ClC at different lipid-to-protein ratios.