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- W2057500296 abstract "Background: We have generated LQT1 and LQT2 rabbit models by over-expressing the Flag-tagged pore mutant KvLQT1-Y315S or HERG-G628S, in the heart of transgenic rabbits. We undertook a proteomics approach in order to identify KvLQT1 as well as HERG-interacting proteins and differences in protein expression in the hearts of these transgenic rabbits.Methods: Triton X-100 solubilized heart membranes were subjected to immuno-precipitation using anti-FLAG antibody. The FLAG-immunoprecipitated KvLQT1 and HERG-bound complexes were resolved by SDS-PAGE and stained with Commassie blue. Upon resolution, 15 protein bands each corresponding to KvLQT1 and HERG-interacting proteins were obtained. Individual protein bands were destained, excised and digested with trypsin. The resulting peptide mixtures were analyzed by high pressure liquid chromatography coupled with electrospray ionization tandem mass spectrometry. The results from mass spectra were searched against the mouse, rat, and human genomic data base using the SEQUEST software. 2-D DIGE technique in conjunction with proteomic studies was employed to investigate the differences in protein expression in the hearts of transgenic rabbits.Results: Our results revealed that both channels were precipitated. In addition, several unique potential KvLQTI and HERG-interacting proteins along with the previously known KvLQT1-interacting protein such as calmodulin, and HERG-interacting proteins, such as Hsc70 and Hsp90 were detected. Our results also showed that there were four protein bands with varying molecular weights which were common for KvLQT1 and HERG-bound complexes, suggesting potential common KvLQT1 and HERG-interacting proteins. Furthermore, the results of 2D-DIGE have revealed the upregulation of several proteins in KvLQT1 transgenic rabbit heart.Conclusions: We have identified potential unique proteins that interact with either KvLQT1 or HERG, as well as the several potential proteins that interact with both of these channels. These interacting proteins might play an important role in regulating the trafficking and localization of these channels." @default.
- W2057500296 created "2016-06-24" @default.
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- W2057500296 date "2009-02-01" @default.
- W2057500296 modified "2023-09-30" @default.
- W2057500296 title "Proteomic Analysis of KvLQT1 and HERG-associated Proteins" @default.
- W2057500296 cites W1992013906 @default.
- W2057500296 doi "https://doi.org/10.1016/j.bpj.2008.12.287" @default.
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