LIPID CONTROL OF G PROTEIN GATED K CHANNEL ACTIVITY

G 蛋白门控 K 通道活性的脂质控制

• 批准号: 6184414
• 负责人: Diomedes E. Logothetis
• 金额: $ 28.11万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-04-01 至 2002-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6184414
• 关键词: G protein; Xenopus; Xenopus oocyte; arrhythmia; chick embryo; enzyme mechanism; ionic bond; membrane activity; phosphatidylinositols; phospholipids; potassium channel; protein structure function; single cell analysis; site directed mutagenesis; sodium potassium exchanging ATPase; voltage /patch clamp; western blottings

Molecular studies of ion channel function have primarily focused on the proteins themselves or on their interactions with other proteins or ions. G protein-gated K channels (KG) for example are thought to be directly activated by the betagamma subunits of GTP binding proteins (Gbetagamma subunits). Current models of KG channel activation involve G protein subunit separation (which renders them active) and interaction with the channel subunits. Our preliminary results suggest that the Gbetagamma subunit/KG channel interaction requires the presence of PIP2 in the membrane in order to manifest its effects on channel activity. This surprising result is accompanied by other effects directly attributable to PIP2, such as the MgATP-dependent sensitization of KG channels to gating by internal Na ions and possibly the MgATP-dependent rundown of G protein stimulation of KG channel activity. These results together with two recent reports on the related inwardly rectifying channel KATP and on the Na / Ca transporter (but not on Na channels or Na / K pumps) herald the potential of an unexplored area of research, crucial to the functional integrity of membrane proteins. Our proposal aims to study in detail the effects of lipids, and in particular phospholipids on KG channel function. The experiments outlined will test further the molecular basis and significance of the PIP2 effects on KG channel activity and the dependence of G protein subunit KG channel activation on the presence of PIP2. It has been proposed that the lipid effects are electrostatic in nature. We will test this hypothesis and seek to identify the basic residues in the channel sequence constituting sites of interaction with the anionic phospholipids. Phospholipids of the phosphoinositide cycle allow dynamic participation of lipids in signaling. We believe that a better appreciation of the molecular details of KG channel function afforded by this study will allow more successful manipulation of this atrial channel in the control of supraventricular arrhythmias. For example, our recent discovery of the MgATP-dependent sensitization of the atrial KG channel (KACh) to gating by internal Na ions allowed us to demonstrate that digitalis treatment causes atrial cells to activate KACh (due to the Na accumulation it causes), providing an important link to the long known effects of this drug on supraventricular rhythm.

离子通道功能的分子研究主要集中在 蛋白质本身或它们与其他蛋白质的相互作用或 离子。例如，G蛋白门控K通道(KG)被认为是 由GTP结合蛋白的β-亚基直接激活 (Gbetagamma亚基)。目前KG通道激活的模型包括 G蛋白亚基分离(使它们变得活跃)和相互作用 具有通道子单元。我们的初步结果表明， Gbetagamma亚基/KG通道相互作用需要PIP2的存在 以显示其对通道活性的影响。 这一令人惊讶的结果还直接伴随着其他影响 归因于PIP2，如KG的镁-三磷酸腺苷依赖的敏化 内钠离子和可能的镁-三磷酸腺苷依赖的门控通道 抑制G蛋白对KG通道活性的刺激。这些结果 以及最近关于相关内向整顿的两篇报道 通道KATP和钠/钙转运体(但不在钠通道或 Na/K泵)预示着一个未被探索的研究领域的潜力， 对膜蛋白的功能完整性至关重要。我们的建议 旨在详细研究脂质的影响，特别是 磷脂对KG通道功能的影响。概述的实验将 进一步验证PIP2效应的分子基础和意义 KG通道活性与G蛋白亚基KG依赖性的研究 在存在PIP2的情况下激活通道。有人提议， 脂类的作用本质上是静电作用。我们将对此进行测试 假设并寻求确定通道中的基本残基 构成与阴离子相互作用部位的序列 磷脂。磷脂酰肌醇循环的磷脂允许 脂质在信号转导中的动态参与。我们相信，一个更好的 KG通道功能的分子细节研究进展 通过这项研究将允许更成功地操作这一心房 经络在控制室上性心律失常中的作用。例如, 我们最近发现的依赖于镁三磷酸腺苷的心房敏化 公斤通道(KACH)到门控由内部的钠离子允许我们 证明洋地黄治疗可使心房细胞激活 卡赫(由于它造成的钠积累)，提供了一个重要的联系 这种药物对室上性心律的作用由来已久。