Phospho-dependent binding of the clathrin AP2 adaptor complex to GABAA receptors regulates the efficacy of inhibitory synaptic transmission

被引:138
作者
Kittler, JT
Chen, GJ
Honing, S
Bogdanov, Y
McAinsh, K
Arancibia-Carcamo, IL
Jovanovic, JN
Pangalos, MN
Haucke, V
Yan, Z
Moss, SJ
机构
[1] Free Univ Berlin, Inst Chem Biochem, D-14195 Berlin, Germany
[2] Wyeth Ayerst Res, Neurosci Discovery, Princeton, NJ 08552 USA
[3] Univ London, Sch Pharm, Dept Pharmacol, London WC1N 1AX, England
[4] UCL, Dept Pharmacol, London WC1E 6BT, England
[5] Univ Penn, Dept Neurosci, Philadelphia, PA 19104 USA
[6] Univ Gottingen, Inst Biochem 2, D-37073 Gottingen, Germany
[7] SUNY Buffalo, Sch Med & Biomed Sci, Dept Physiol & Biophys, Buffalo, NY 14214 USA
[8] UCL, Dept Physiol, London WC1E 6BT, England
基金
英国惠康基金;
关键词
endocytosis; phosphorylation;
D O I
10.1073/pnas.0506653102
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The efficacy of synaptic inhibition depends on the number of gamma-aminobutyric acid type A receptors (GABA(A)Rs) expressed on the cell surface of neurons. The clathrin adaptor protein 2 (AP2) complex is a critical regulator of GABA(A)R endocytosis and, hence, surface receptor number. Here, we identify a previously uncharacterized atypical AP2 binding motif conserved within the intracellular domains of all GABA(A)R beta subunit isoforms. This AP2 binding motif (KTHLRRRSSQLK in the beta 3 subunit) incorporates the major sites of serine phosphorylation within receptor beta subunits, and phosphorylation within this site inhibits AP2 binding. Furthermore, by using surface plasmon resonance, we establish that a peptide (pep beta 3) corresponding to the AP2 binding motif in the GABA(A)R beta 3 subunit binds to AP2 with high affinity only when dephosphorylated. Moreover, the pep beta 3 peptide, but not its phosphorylated equivalent (pep beta 3-phos), enhanced the amplitude of miniature inhibitory synaptic current and whole cell GABA(A)R current. These effects of pep beta 3 on GABA(A)R current were occluded by inhibitors of dynamin-dependent endocytosis supporting an action of pep beta 3 on GABA(A)R endocytosis. Therefore phosphodependent regulation of AP2 binding to GABA(A)Rs provides a mechanism to specify receptor cell surface number and the efficacy of inhibitory synaptic transmission.
引用
收藏
页码:14871 / 14876
页数:6
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