The DOI matches its Crossref record; 43 of the 43 references it cites were found in a registry or an index.
Purification of Recombinant G Proteins from Sf9 Cells by Hexahistidine Tagging of Associated Subunits
Open accessAbstract
A method is described for purification of G protein alpha and beta gamma subunits from Sf9 cells infected with recombinant baculoviruses. The subunit to be purified is coexpressed with an associated subunit bearing a hexahistidine tag. After adsorption of the oligomer to a Ni(2+)-containing column, the subunit to be purified is eluted specifically by promoting subunit dissociation with AIF4-. The alpha subunits of G12, Gq, Gz, and Gi1 and the beta 1 gamma 2 subunit complex were easily and efficiently purified by this method. Results was superior to established procedures in all cases. Purified alpha 12 was characterized for the first time. The protein has a slow rate of guanine nucleotide exchange (kon, GTP gamma S = 0.01 min-1) and a very slow kcat for hydrolysis of GTP (0.1-0.2 min-1). GTP gamma S (guanosine 5' -3-O- (thio)triphosphate) alpha 12 does not influence the activity of several adenylyl cyclases or phospholipases. Activated alpha z inhibits the activity of type I and type V adenylyl cyclases. It is a somewhat more potent inhibitor of type V adenylyl cyclase than is activated alpha i1.
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Metadata
- Authors
- Kozasa, T., & Gilman, A. G.
- Venue
- Journal of Biological Chemistry
- Publisher
- Elsevier BV
- Published
- 1995-01
- Volume
- 270
- Issue
- 4
- Pages
- 1734-1741
- ISSN
- 0021-9258 (ISSN-L)
- OpenAlex
- W2148310842
- Cited by
- 334
- References
- 43
- FWCI
- 8.53
- Type
- journal article
- Language
- en
- Open access
- hybrid
- Licence
- www.elsevier.com/tdm/userlicense/1.0/ (tdm)
- creativecommons.org/licenses/by/4.0/ (vor, 9427 d)
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- Kozasa, T., & Gilman, A. G. (1995). Purification of Recombinant G Proteins from Sf9 Cells by Hexahistidine Tagging of Associated Subunits. Journal of Biological Chemistry. https://doi.org/10.1074/jbc.270.4.1734
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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