Roles of G proteins in coupling of receptors to ionic channels and other effector systems.
Guanine nucleotide binding (G) proteins are heterotrimers that couple a wide range of receptors to ionic channels. The coupling may be indirect, via cytoplasmic agents, or direct, as has been shown for two K+ channels and two Ca2+ channels. One example of direct G protein gating is the atrial muscarinic K+ channel K+[ACh], an inwardly rectifying K+ channel with a slope conductance of 40 pS in symmetrical isotonic K+ solutions and a mean open lifetime of 1.4 ms at potentials between -40 and -100 mV. Another is the clonal GH3 muscarinic or somatostatin K+ channel, also inwardly rectifying but with a slope conductance of 55 pS. A G protein, Gk, purified from human red blood cells (hRBC) activates K+ [ACh] channels at subpicomolar concentrations; its alpha subunit is equipotent. Except for being irreversible, their effects on gating precisely mimic physiological gating produced by muscarinic agonists. The alpha k effects are general and are similar in atria from adult guinea pig, neonatal rat, and chick embryo. The hydrophilic beta gamma from transducin has no effect while hydrophobic beta gamma from brain, hRBCs, or retina has effects at nanomolar concentrations which in our hands cannot be dissociated from detergent effects. An anti-alpha k monoclonal antibody blocks muscarinic activation, supporting the concept that the physiological mediator is the alpha subunit not the beta gamma dimer. The techniques of molecular biology are now being used to specify G protein gating. A "bacterial" alpha i-3 expressed in Escherichia coli using a pT7 expression system mimics the gating produced by hRBC alpha k.
Birnbaumer
L
L
Department of Cell Biology, Baylor College of Medicine, Houston, Texas.
Abramowitz
J
J
Yatani
A
A
Okabe
K
K
Mattera
R
R
Graf
R
R
Sanford
J
J
Codina
J
J
Brown
A M
AM
D000818:Animals; D018407:Bacterial Physiological Phenomena; D015220:Calcium Channels / Q000502:physiology; D019204:GTP-Binding Proteins / Q000502:physiology*; D015640:Ion Channel Gating; D015221:Potassium Channels / Q000502:physiology*; D015398:Signal Transduction
D015220:Calcium Channels; D015221:Potassium Channels; D019204:GTP-Binding Proteins