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Published on: March 9, 2012
Transglutaminase-catalyzed transamidation: a novel mechanism for Rac1 activation by 5-hydroxytryptamine2A receptor
1Neuroscience Program, Loyola University Medical Center, Maywood, Illinois, USA.
Serotonin activates Rac1 in neurons through transglutaminase (TGase) and 5-hydroxytryptamine (HT)(2A) receptors. This novel signaling pathway involves TGase-catalyzed serotonin binding to Rac1, transiently increasing its activity.
Area of Science:
- Neuroscience
- Molecular Biology
- Biochemistry
Background:
- Platelet aggregation is mediated by transglutaminase (TGase) activation of small G proteins via 5-hydroxytryptamine (HT)(2A) receptors.
- The role of TGase and 5-HT(2A) receptors in neuronal signaling remains to be fully elucidated.
Purpose of the Study:
- To investigate the hypothesis that 5-HT(2A) receptor stimulation in neurons activates TGase, leading to Rac1 transamidation and activation.
- To identify the molecular mechanism of 5-HT(2A) receptor signaling in neurons involving TGase and Rac1.
Main Methods:
- Immunoprecipitation and immunoblotting in rat cortical cell line A1A1v.
- Treatment with selective 5-HT receptor agonists and antagonists.
- Inhibition of TGase using cystamine and small interfering RNA.
- Coimmunoprecipitation experiments to detect serotonin binding to Rac1.
Main Results:
- Serotonin stimulation of 5-HT(2A) receptors increased TGase-catalyzed transamidation of serotonin to Rac1 in neuronal cells.
- This modification and subsequent Rac1 activation were transient, peaking at 5-15 minutes.
- Inhibition of TGase blocked Rac1 activation and serotonin binding to Rac1.
Conclusions:
- TGase-catalyzed transamidation of serotonin to Rac1, mediated by 5-HT(2A) receptor stimulation, is a novel mechanism for transient Rac1 activation in neurons.
- This pathway represents a new effector and second messenger in the 5-HT(2A) receptor signaling cascade.
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