Summary
The purification of recombinant G protein a subunits expressed in Escherichia coli (E. coli) is a convenient and inexpensive method to obtain homogeneous preparations of protein for biochemical and biophysical analyses. Wild-type and mutant forms of Gα are easily produced for analysis of their intrinsic biochemical properties, as well as for reconstitution with receptors, effectors, regulators, and G protein βγ subunits. Methods are described for the expression of Giα and Gsα proteins in E. coli. Protocols are provided for the purification of untagged G protein a subunits using conventional chromatography and histidine (His)-tagged subunits using metal chelate chromatography. Modification of Gα with myristate can be recapitulated in E. coli by expressing N-myristoyltransferase (NMT) with its G protein substrate. Protocols for the production and purification of myristoylated Gα are presented.
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Greentree, W.K., Linder, M.E. (2004). Purification of Recombinant G Protein α Subunits from Escherichia coli . In: Smrcka, A.V. (eds) G Protein Signaling. Methods in Molecular Biology™, vol 237. Humana Press. https://doi.org/10.1385/1-59259-430-1:3
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DOI: https://doi.org/10.1385/1-59259-430-1:3
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