Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
9954234 | Protein Expression and Purification | 2019 | 31 Pages |
Abstract
The main principles of higher-order protein oligomerization are elucidated by many structural and biophysical studies. An astonishing number of proteins self-associate to form dimers or higher-order quaternary structures which further interact with other biomolecules to elicit complex cellular responses. In this study, we describe a simple and convenient approach to determine the oligomeric state of purified protein complexes that combines implementation of a novel form of clear-native gel electrophoresis and size exclusion chromatography in line with multi-angle light scattering. Here, we demonstrate the accuracy of this ensemble approach by characterizing the previously established pentameric state of the intracellular domain of serotonin type 3A (5-HT3A) receptors.
Keywords
BN-PAGECN-PAGESEC-MALS5-HT3ApLGICDeoxyribonuclease I6-AHA6-aminohexanoic acidE. coliPMSFIPTGDNase IMBPBSAProtein oligomerizationtris(2-carboxyethyl)phosphine hydrochloridebovine serum albuminEscherichia coliNative gel electrophoresisblue-native polyacrylamide gel electrophoresisisopropyl β-D-1-thiogalactopyranosideintracellular domainICDphenylmethylsulfonyl fluoridemulti-angle light scatteringmaltose-binding proteinPentameric ligand-gated ion channelSize exclusion chromatography
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Authors
Akash Pandhare, Antonia G. Stuebler, Elham Pirayesh, Michaela Jansen,