| Autor: |
Wendy K, Greentree, Maurine E, Linder |
| Rok vydání: |
2003 |
| Předmět: |
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| Zdroj: |
Methods in molecular biology (Clifton, N.J.). 237 |
| ISSN: |
1064-3745 |
| Popis: |
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 G alpha are easily produced for analysis of their intrinsic biochemical properties, as well as for reconstitution with receptors, effectors, regulators, and G protein beta gamma subunits. Methods are described for the expression of Gi alpha and Gs alpha 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 G alpha 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 G alpha are presented. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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