A point mutation in the ectodomain-transmembrane 2 interface eliminates the inhibitory effects of ethanol in P2X4 receptors.
A point mutation in the ectodomain-transmembrane 2 interface eliminates the inhibitory effects of ethanol in P2X4 receptors.
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DOI：
10.1111/j.1471-4159.2009.06460.x
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发表时间：
2010-01
影响因子：
4.7
通讯作者：
Davies DL
中科院分区：
文献类型：
--
作者：
Popova M;Asatryan L;Ostrovskaya O;Wyatt LR;Li K;Alkana RL;Davies DL
ATP-gated P2X4 receptors (P2X4R) are abundantly expressed in the CNS. However, little is known about the molecular targets for ethanol action in P2X4Rs. The current investigation tested the hypothesis that the ectodomain-TM interface contains residues that are important for the action of ethanol in P2X4Rs. Wildtype (WT) and mutant P2X4R were expressed in Xenopus oocytes. ATP concentration-response curves and ethanol (10–200 mM)-induced changes in ATP EC10-gated currents were determined using two-electrode voltage clamp (−70 mV). Alanine substitution at the ectodomain-TM1 interface (positions 50–61) resulted in minimal changes in ethanol response. On the other hand, alanine substitution at the ectodomain-TM2 interface (positions 321–337) identified two key residues (D331 and M336) that the alanine mutation significantly reduced ethanol inhibition of ATP-gated currents without causing marked changes in ATP Imax, EC50 or Hill slope. Other amino acid substitutions at positions 331 and 336 significantly altered or eliminated the modulatory effects of ethanol. Linear regression analyses revealed a significant relationship between hydropathy and polarity, but not molecular volume/molecular weight of the residues at these two positions. The results support the proposed hypothesis and represent an important step towards developing ethanol-insensitive receptors for investigating the role of P2X4Rs in mediating behavioral effects of ethanol.
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影响因子：
11.4
作者：
Buell, G;Lewis, C;Surprenant, A
通讯作者：
Surprenant, A
影响因子：
4.8
作者：
Perkins, Daya I.;Trudell, James R.;Davies, Daryl L.
通讯作者：
Davies, Daryl L.
影响因子：
4.7
作者：
Crawford, Daniel K.;Trudell, James R.;Alkana, Ronald L.
通讯作者：
Alkana, Ronald L.
影响因子：
4.7
作者：
Asatryan, Liana;Popova, Maya;Davies, Daryl L.
通讯作者：
Davies, Daryl L.
影响因子：
5
作者：
Davies, DL;Alkana, RL
通讯作者：
Alkana, RL