2010م - 1444هـ
نبذه عن الكتاب:



Purification of Recombinant G Protein α Subunits

from Escherichia coli

Wendy K. Greentree and Maurine E. Linder

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 Gi

α 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.

Key Words: G protein; α subunit; signal transduction; protein purification; affinity chromatography; GTPase; membrane protein; myristoylation;

N-myristoyltransferase.