PKC MODULATION OF GS8 AND CALCIUM CHANNEL INTERACTIONS

GS8 和钙通道相互作用的 PKC 调节

• 批准号: 6185616
• 负责人: KAISA H HELLEVUO
• 金额: $ 5.88万
• 依托单位: UNIVERSITY OF COLORADO DENVER
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2000-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6185616
• 关键词: actin binding protein; biological signal transduction; calcium channel; cell line; electrophysiology; enzyme mechanism; enzyme substrate; isozymes; opioid receptor; phosphorylation; protein isoforms; protein kinase C; receptor coupling; site directed mutagenesis; synthetic peptide; transfection; voltage /patch clamp; voltage gated channel

DESCRIPTION: (Applicant's Abstract) My predoctoral and post- doctoral background consists of behavioral, neurochemical, biochemical and molecular biology studies. During my postdoctoral years under the supervision of Dr. Boris Tabakoff, Department of Pharmacology, UCHSC, I developed a special interest and expertise in G protein-mediated signal transduction, especially in the family of adenylyl. cyclases. Now my goal is to expand my expertise in this area and to acquire skills needed to study how G proteins regulate another class of effectors, the voltagegated calcium channels. In the proposed training I will learn electrophysiology, especially whole cell patch clamp recordings. This technique will allow me to measure events of G protein regulation of the channels in the time scale of milliseconds-seconds, as opposed to the time scale of minutes in the adenylyl cyclase enzyme assays. The proposed training will be take place in Dr. William Sather's (Mentor) laboratory at the Deparunent of Pharmacology, UCHSC. Dr. Sather's laboratory uses routinely electrophysiological techniques, such as whole cell and single channel patch clamp and two-electrode voltage clamp techniques. The Co-Mentor, Dr. Thomas Dunwiddie is another expert electrophysiologist. These laboratories and the Department provide excellent facilities and intellectual milieu. The research project will focus on protein kinase C (PKC) and G protein modulation of N- and P/Q-type calcium channels. Gi/Go- coupled receptors have been demonstrated to inhibit, in a membrane- delimited manner, N- and P/Q-type channels that are involved in neurotransmitter release in the CNS. Recent studies have shown that GBy subunits which are released upon Gi/Go-coupled receptor stimulation are directly interacting with the chafifiel al subunits inhibiting the channel function. On the other hand, there is evidence that activation of PKC antagonizes the membrane-delimited inhibition of calcium channels by GBy. The aim of the proposed studies is to investigate in detail the role of different PKC isozymes and the mechanisms how these isozyntes modulate the interactions of Gpy with N (alB)- and PIQ (aW-type channels. This training will allow me in the future to write proposals and receive funding for studies on G protein regulation of different effector classes as a fully independent investigator, which will advance my career in the academic environment. Calcium channels play a key role in neurons and have been implicated as targets of diverse neurological diseases, such as familial herniplegic migraine, episodic ataxia I type 2 and spinocerebellar ataxia 6.

产品说明：我的博士前和博士后背景包括行为学、神经化学、生物化学和分子生物学研究。在UCHSC药理学系Boris Tabakoff博士的指导下，我对G蛋白介导的信号转导特别感兴趣，特别是在腺苷酸家族中。环化酶现在，我的目标是扩展我在这一领域的专业知识，并获得研究G蛋白如何调节另一类效应器（电压门控钙通道）所需的技能。在建议的培训中，我将学习电生理学，特别是全细胞膜片钳记录。这项技术将允许我在毫秒-秒的时间尺度内测量通道的G蛋白调节事件，而不是腺苷酸环化酶测定中的分钟时间尺度。拟定的培训将在UCHSC药理学实验室的William Sather博士（导师）实验室进行。Sather博士的实验室常规使用电生理技术，如全细胞和单通道膜片钳和双电极电压钳技术。共同导师，博士托马斯邓威迪是另一个专家电生理学家。这些实验室和该部门提供了一流的设施和知识环境。该研究项目将重点关注蛋白激酶C（PKC）和G蛋白对N型和P/Q型钙通道的调节。已经证明Gi/Go偶联受体以膜界定的方式抑制参与CNS中神经递质释放的N型和P/Q型通道。最近的研究表明，在Gi/Go偶联受体刺激时释放的GBy亚基直接与抑制通道功能的chafielal亚基相互作用。另一方面，有证据表明PKC的激活可以拮抗GBy对钙通道的膜界定抑制。本研究的目的是详细研究不同PKC同工酶的作用以及这些同工酶如何调节Gpy与N（aB）-和PIQ（aW）-型通道的相互作用的机制。这项培训将使我在未来写的建议，并获得资金的研究G蛋白调节不同的效应器类作为一个完全独立的调查员，这将推动我的职业生涯在学术环境。钙通道在神经元中起着关键作用，并被认为是多种神经系统疾病的靶点，如家族性疝气性偏头痛、发作性共济失调I型2和脊髓小脑共济失调6。