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- W2912015821 abstract "Several recently-released structures of the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) protein in a detergent-micelle have enabled great progress in understanding CFTR; however, many labs report that CFTR has significantly reduced ATPase activity in detergent-micelles and has reduced stability as exhibited by aggregation over short periods of time. This begs the question, why is wildtype-CFTR (wt-CFTR) so unstable in detergent, and would a lipid environment improve stability? Is the ATPase rate, or channel activity, itself, different when CFTR is solubilized in detergent as compared to when it is purified within the native lipid environment? And what lipids are natively surrounding and interacting with CFTR? To begin answering these questions, we are using a variety of biochemical and biophysical techniques, including the following: detergent-solubilized protein purification, size-exclusion chromatography, reconstitution (into proteoliposomes and nanodiscs), styrene maleic acid lipid particles (SMALP)-based protein purification, ATPase assay, mass-spectrometry lipidomics analysis, and transmission electron microscopy. Results thus far indicate that CFTR function, at least as measured by intrinsic ATPase activity, is much higher when CFTR is embedded in a lipid nanodisc than when surrounded by detergent. These results suggest that CFTR function is indeed dependent upon its environment. Studies are underway to further characterize the lipid-dependence on the activity of CFTR via reconstitution in different lipid mixtures and SMALP purification. Mass-spectrometry lipidomics analysis provides a highly detailed characterization that can be correlated to activity in the different lipid environments. (Supported by CF Fdn. MCCART17G0, CF Fdn. MCCART15R0, and Marcus Professorship). Key words: Cystic Fibrosis Transmembrane conductance Regulator (CFTR), nanodiscs, Styrene Maleic Acid lipid particle (SMALP), lipidomics." @default.
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- W2912015821 date "2019-02-01" @default.
- W2912015821 modified "2023-09-26" @default.
- W2912015821 title "Effects of Lipid and Detergent Environments on Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) Function and Structure" @default.
- W2912015821 doi "https://doi.org/10.1016/j.bpj.2018.11.1222" @default.
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