Related Experiment Video
Updated: Oct 4, 2026

Comparing the Affinity of GTPase-binding Proteins using Competition Assays
Published on: October 8, 2015
An intramolecular contact in Galpha transducin that participates in maintaining its intrinsic GDP release rate
1Institute for Neuroscience, Northwestern University, Chicago, Illinois, USA.
Abstract:
Receptor mediated stimulation of the G protein-alpha subunit leads to exchange of GDP for GTP, activating the protein. Spontaneous GDP release from Galpha can also lead to the active state, if GTP in solution binds the nucleotide binding pocket. The purpose of this study is to evaluate the molecular determinants for maintaining the spontaneous GDP release rates between two Galpha subunits. Galpha(t) has a low rate of nucleotide release, compared to Galpha(i1). Galpha(t/i1) chimeras were used to explore the molecular basis for this behavior. The C-terminal alpha4-helix, the N-terminal 56 residues and the Switch I/II regions of Galpha(t) were shown to affect the low spontaneous GDP release rate in Galpha(t). A specific molecular contact between Asp26 and Asn191 was found in Galpha(t) that is not present in Galpha(i1). In two chimeras disrupting this interaction produced an increased spontaneous GDP release; restoring the contact present in Galpha(t) into these chimeras decreased the GDP release rate by half as compared to the original chimeras. Similarly, introduction of this contact in wild-type Galpha(i1) decreased the GDP release rate of Galpha(i1) by half. Differences in GDP release rates may reflect physiological roles these proteins play in living systems.
Related Concept Videos
Activation and Inactivation of G Proteins
GTPases and their Regulation
Large G-proteins, also known...
GTPases and their Regulation
Large G-proteins, also known...
G Protein-coupled Receptors
G-protein Coupled Receptors
G Protein-coupled Receptors
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...

