Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
2021063 | Protein Expression and Purification | 2011 | 6 Pages |
Abstract
Previous studies of constitutively activated mutants of opsin in the absence of chromophore were carried out in crude cell membranes because such mutants could not be recovered in a detergent-solubilized form in the active state. We employed a strategy in which a stabilizing disulfide bond allowed for successful purification of a constitutively activated mutant opsin, N2C/E113Q/M257Y/D282C, solubilized in nonionic detergent from mammalian cell culture. The purified mutant opsin is able to activate transducin to a higher degree than opsin and may prove useful for future structural studies of the active state of GPCRs.
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Authors
K. Christopher Min, Yan Jin, Wayne A. Hendrickson,