Abstract
Purified guanine nucleotide-binding regulatory proteins, as either the oligomers or the isolated nucleotide-binding α subunits, display anomalous kinetics of nucleotide binding. This is due to the presence of tightly bound GDP in these preparations. The dissociation of bound GDP is the rate-limiting step for nucleotide binding. GDP can be removed by chromatography in the presence of 1 M (NH4)2SO4 and 20% glycerol, which yields preparations of G proteins that contain less than 0.1 mol of GDP/mol of guanosine 5'-(γ-thio)triphosphate (GTPγS)-binding site. When the GDP is removed, the binding of GTPγS displays kinetics consistent with a bimolecular reaction.
Original language | English (US) |
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Pages (from-to) | 7393-7399 |
Number of pages | 7 |
Journal | Journal of Biological Chemistry |
Volume | 261 |
Issue number | 16 |
State | Published - 1986 |
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology
- Cell Biology