Evolutionary Biophysics of Slo3 Potassium Channels

Slo3 钾通道的进化生物物理学

基本信息

批准号：
9062306
负责人：
Benjamin J Liebeskind
金额：
$ 5.43万
依托单位：
UNIVERSITY OF TEXAS AT AUSTIN
依托单位国家：
美国
项目类别：
财政年份：
2015
资助国家：
美国
起止时间：
2015-05-01 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9062306
关键词：
Accounting Achievement Affect Animal Model Belief Binding Binding Sites Biology Biophysics Birds Calcium Calcium Binding Cells Cellular Membrane Chimera organism DNA Sequence Data Analyses Discipline Disease Drug Targeting Electrophysiology (science)Elements Epilepsy Evolution Family Future Genes Genome Histidine Homologous Gene Human Ion Channel Ions Knowledge Laboratories Labyrinth Learning Male Contraceptive Agents Mammals Medical Membrane Modeling Molecular Molecular Evolution Movement Mus Muscle Contraction Mutation Nervous system structure Pharmaceutical Preparations Phylogenetic Analysis Physiology Play Potassium Potassium Channel Proteins Protons Recording of previous events Reporting Reproductive Biology Reptiles Research Role Scientist Site Site-Directed Mutagenesis Smooth Muscle Sperm Capacitation Statistical Models Structure Techniques Testing Time Tissues Training Work base comparative comparative genomics electrical potential large-conductance calcium-activated potassium channels novel novel strategies potassium ion public health relevance reconstruction relating to nervous system sensor single molecule sperm cell sperm function targeted treatment theories voltage

项目摘要

DESCRIPTION (provided by applicant): Potassium ions are used by the body to build up an electrical potential across cellular membranes. This potential is used as energy to drive the movement of molecules into and out of cells, and, most famously, to drive the electrical activity that characterizes the nervous system. Potassium channels are the proteins that allow these important ions across the membrane, and their structure and function are well known from 60 years of biophysical and structural research. One notable group of potassium channels, known as Slo channels, opens and closes depending on both the membrane voltage and the concentration of various intracellular ions. These channels are involved in diverse elements of physiology, including the timing of neural activity, the function of the inner ear, smooth muscle contraction, and sperm capacitation. Mutations in Slo channels are associated with epilepsy, and they are potential drug targets for a variety of medical purposes. In the Slo family, the best understand channel, Slo1, also called the BK channel, is closely related to the least well understood channel, Slo3. Despite being closely related, the two channel types are affected by different intracellular ions and are expressed in different tissues. Slo1 is sensitive to intracelllar calcium, while Slo3 appears to be primarily sensitive to pH. Slo3 has primarily been characterized in mice, however, and current research suggests human Slo3 might be very different from mice. There are also many outstanding questions about the locus of proton binding in Slo3. My proposed work uses a different approach than the one taken so far in Slo3 research. Rather than look in a few model organisms, I propose to take a comparative and evolutionary approach that can reconstruct the story of how Slo3 came to be different from Slo1. I have used comparative genomics and phylogenetics to learn new things about Slo3 and generate predictive hypotheses about the molecular basis of its function. Contrary to the current belief that Slo3 exists only in mammals, I have found Slo3 genes in the genomes of birds and reptiles. To reconstruct the history of Slo3, I predicted the sequence of the ancestors of extant Slo3 channels using statistical models of evolution. These new channels, extant and ancestral, tell a predictive story of how Slo3 differentiated from Slo1 by losing known calcium binding sites and acquiring new sites that may sense intracellular pH. My proposed work will test these hypotheses using heterologous expression and biophysics. The work will be done in the laboratory of Dr. Richard Aldrich, who has contributed much of the current knowledge of Slo channel function.

描述（由申请人提供）：身体利用钾离子在细胞膜上建立电势，该电势被用作驱动分子进出细胞的运动的能量，最著名的是驱动细胞的运动。钾通道是神经系统的电活动特征，它是允许这些重要离子穿过细胞膜的蛋白质，其结构和功能在 60 年的生物物理和结构研究中已众所周知。一组著名的钾通道，称为 Slo。这些通道的打开和关闭取决于膜电压和各种细胞内离子的浓度，这些通道涉及多种生理学要素，包括神经活动的时间、内耳的功能、平滑肌收缩和精子获能。 Slo 通道的突变与癫痫有关，它们是多种医学用途的潜在药物靶点。最了解的通道 Slo1，也称为 BK 通道，与最不了解的通道 Slo3 密切相关，尽管密切相关，但这两种通道类型受不同的细胞内离子影响，并且在不同的组织中表达。然而，Slo3 似乎主要对 pH 敏感，目前的研究表明人类 Slo3 可能与小鼠有很大不同。关于质子结合位点也存在许多悬而未决的问题。 Slo3。我提出的工作采用了与 Slo3 研究迄今为止所采用的方法不同的方法，我建议采用一种比较和进化的方法来重建 Slo3 与 Slo3 的不同之处。我使用比较基因组学和系统发育学来了解有关 Slo3 的新知识，并对其功能的分子基础提出预测假设，与目前认为 Slo3 仅存在于哺乳动物中的观点相反，我在哺乳动物中发现了 Slo3 基因。为了重建 Slo3 的历史，我使用进化统计模型预测了现存 Slo3 通道的祖先序列，这些新通道（现存的和祖先的）讲述了 Slo3 如何通过丢失而与 Slo1 区分开来的预测故事。我提议的工作将利用异源表达和生物物理学来测试这些假设，该工作将在 Richard Aldrich 博士的实验室中完成。贡献了当前 Slo 通道函数的大部分知识。