Huynh Thuan G, Cuny Hartmut, Slesinger Paul A, Adams David J
Health Innovations Research Institute, RMIT University, Melbourne, Victoria, Australia (T.G.H., H.C., D.J.A.); and Peptide Biology Laboratories, The Salk Institute for Biological Studies, La Jolla, California (P.A.S.).
Mol Pharmacol. 2015 Feb;87(2):240-50. doi: 10.1124/mol.114.096156. Epub 2014 Nov 25.
Neuronal voltage-gated N-type (Cav2.2) calcium channels are expressed throughout the nervous system and regulate neurotransmitter release and hence synaptic transmission. They are predominantly modulated via G protein-coupled receptor activated pathways, and the well characterized Gβγ subunits inhibit Cav2.2 currents. Analgesic α-conotoxin Vc1.1, a peptide from predatory marine cone snail venom, inhibits Cav2.2 channels by activating pertussis toxin-sensitive Gi/o proteins via the GABAB receptor (GABA(B)R) and potently suppresses pain in rat models. Using a heterologous GABA(B)R expression system, electrophysiology, and mutagenesis, we showed α-conotoxin Vc1.1 modulates Cav2.2 via a different pathway from that of the GABA(B)R agonists GABA and baclofen. In contrast to GABA and baclofen, Vc1.1 changes Cav2.2 channel kinetics by increasing the rate of activation and shifting its half-maximum inactivation to a more hyperpolarized potential. We then systematically truncated the GABA(B)(1a) C terminus and discovered that removing the proximal carboxyl terminus of the GABA(B)(1a) subunit significantly reduced Vc1.1 inhibition of Cav2.2 currents. We propose a novel mechanism by which Vc1.1 activates GABA(B)R and requires the GABA(B)(1a) proximal carboxyl terminus domain to inhibit Cav2.2 channels. These findings provide important insights into how GABA(B)Rs mediate Cav2.2 channel inhibition and alter nociceptive transmission.
神经元电压门控N型(Cav2.2)钙通道在整个神经系统中表达,并调节神经递质释放,进而调节突触传递。它们主要通过G蛋白偶联受体激活途径进行调节,且已被充分表征的Gβγ亚基可抑制Cav2.2电流。镇痛性α-芋螺毒素Vc1.1是一种来自掠食性海洋芋螺毒液的肽,它通过GABAB受体(GABA(B)R)激活百日咳毒素敏感的Gi/o蛋白来抑制Cav2.2通道,并在大鼠模型中有效抑制疼痛。利用异源GABA(B)R表达系统、电生理学和诱变技术,我们发现α-芋螺毒素Vc1.1通过一条与GABA(B)R激动剂GABA和巴氯芬不同的途径调节Cav2.2。与GABA和巴氯芬不同,Vc1.1通过增加激活速率并将其半数最大失活电位移至更超极化的电位来改变Cav2.2通道动力学。然后,我们系统地截短了GABA(B)(1a)的C末端,发现去除GABA(B)(1a)亚基的近端羧基末端可显著降低Vc1.1对Cav2.2电流的抑制作用。我们提出了一种新机制,即Vc1.1激活GABA(B)R并需要GABA(B)(1a)近端羧基末端结构域来抑制Cav2.2通道。这些发现为GABA(B)R如何介导Cav2.2通道抑制并改变伤害性传递提供了重要见解。