Related Experiment Videos
Phosphorylation of the nicotinic acetylcholine receptor regulates its rate of desensitization
Abstract:
Recent studies have provided evidence for a role of protein phosphorylation in the regulation of the function of various potassium and calcium channels (for reviews, see refs 1, 2). As these ion channels have not yet been isolated and characterized, it has not been possible to determine whether phosphorylation of the ion channels themselves alters their properties or whether some indirect mechanism is involved. In contrast, the nicotinic acetylcholine receptor, a neurotransmitter-dependent ion channel, has been extensively characterized biochemically and has been shown to be directly phosphorylated. The phosphorylation of this receptor is catalysed by at least three different protein kinases (cyclic AMP-dependent protein kinase, protein kinase C and a tyrosine-specific protein kinase) on seven different phosphorylation sites. However, the functional significance of phosphorylation of the receptor has been unclear. We have now examined the functional effects of phosphorylation of the nicotinic acetylcholine receptor by cAMP-dependent protein kinase. We investigated the ion transport properties of the purified and reconstituted acetylcholine receptor before and after phosphorylation. We report here that phosphorylation of the nicotinic acetylcholine receptor on the gamma- and delta-subunits by cAMP-dependent protein kinase increases the rate of the rapid desensitization of the receptor, a process by which the receptor is inactivated in the presence of acetylcholine (ACh). These results provide the first direct evidence that phosphorylation of an ion channel protein modulates its function and suggest that phosphorylation of postsynaptic receptors in general may play an important role in synaptic plasticity.
Insights
Protein phosphorylation directly impacts ion channel function. Phosphorylation of the nicotinic acetylcholine receptor by cAMP-dependent protein kinase accelerates its desensitization, suggesting a role in synaptic plasticity.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Protein phosphorylation regulates ion channel function, but direct evidence is limited for channels like potassium and calcium.
- The nicotinic acetylcholine receptor (nAChR) is a neurotransmitter-dependent ion channel extensively characterized and known to be directly phosphorylated.
- The functional impact of nAChR phosphorylation by various kinases remains unclear.
Purpose of the Study:
- To investigate the functional consequences of direct phosphorylation of the nicotinic acetylcholine receptor by cAMP-dependent protein kinase.
- To determine if phosphorylation of the nAChR protein alters its ion transport properties.
Main Methods:
- Purification and reconstitution of the nicotinic acetylcholine receptor.
- Comparative analysis of ion transport properties before and after phosphorylation by cAMP-dependent protein kinase.
Main Results:
- Phosphorylation of the gamma- and delta-subunits of the nAChR by cAMP-dependent protein kinase was observed.
- This phosphorylation significantly increases the rate of rapid desensitization of the receptor in the presence of acetylcholine (ACh).
Conclusions:
- Provides the first direct evidence that phosphorylation of an ion channel protein modulates its function.
- Suggests that phosphorylation of postsynaptic receptors may be crucial for synaptic plasticity.