Principles:: Extending the utility of [35S]GTPγS binding assays

Ligand regulation of the binding of [S-35]GTPgammaS is one of the most widely used methods to measure receptor activation of heterotrimeric G proteins. However, until recently this method was largely restricted to receptors that interact with members of the family of pertussis-toxin-sensitive G proteins. Here, the reasons for this restriction are discussed and recent approaches that have extended the utility of this method such that it is now suitable for analysis of the activation of any heterotrimeric G protein are reviewed.