Calcium desensitization in Smooth Muscle

平滑肌钙脱敏

基本信息

批准号：
7388268
负责人：
TIMOTHY A HAYSTEAD
金额：
$ 30.43万
依托单位：
DUKE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2006
资助国家：
美国
起止时间：
2006-04-01 至 2010-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7388268
关键词：
Adrenergic Agents Adult Affect Affinity Agonist Amino Acids Binding Blood Pressure Calcium Cardiovascular Physiology Cell Surface Receptors Cell membrane Cells Characteristics Cholinergic Agents Contracts Cyclic AMP Cyclic GMP Cyclic Nucleotides Disease Electron Microscopy Elements Enzymes Event Family Fluorescence Gastrointestinal Diseases Genes Genetic Glaucoma Histocytochemistry Hormones Human Human Genome Hypertension Immune Individual Knockout Mice Lead Mass Spectrum Analysis Mechanics Mediating Molecular Molecular Biology Mus Muscle Muscle Fibers Muscle Proteins Muscle function Muscle relaxation phase Neurons Neurotransmitters Organelles Pattern Pharmaceutical Preparations Phenotype Phosphoproteins Phosphorylation Phosphorylation Site Physiological Physiological Processes Physiology Play Process Property Protein Kinase Proteins Proteomics Purpose Regulation Relaxation Research Personnel Respiration Role Sexual Dysfunction Signal Transduction Signal Transduction Pathway Site-Directed Mutagenesis Smooth Muscle Spasm Striated Muscles Structure Techniques Testing Thick Thin Filament Thinking Tissues adrenergic base cell motility cell type cholinergic desensitization gastrointestinal human disease in vivo mouse genome nerve supply novel programs reproductive function research study response

项目摘要

Smooth muscle plays an essential role in a wide variety of physiological processes, and although the basic function of every smooth muscle is the same, to contract and relax, the mechanical properties and responsiveness to hormones, neurotransmitters and drugs varies greatly between smooth muscle types. Factors that dictate the contractile characteristics of smooth muscle include plasma membrane properties, ratio and compliment of signal transducing proteins, the composition of the contractile apparatus itself. Alteration in the normal blend of these components is thought to underlie the molecular basis of several human diseases that involve smooth muscle including hypertension, bronco spasm, sexual dysfunction, gastrointestinal disorders and glaucoma. It is our hypothesis that by studying the molecular processes by which individual smooth muscles normally respond to stimulation will lead to more selective therapies to treat these disorders. The recent completion of the human and mouse genomes in combination with advanced techniques in mass spectrometry affords new opportunities for probing signal transduction pathways in cells. In this proposal we will employ a unique combination of proteomics, muscle physiology, molecular biology, immuno-histochemistry and mouse genetics to determine the molecular mechanisms by which cGMP through the activation of cyclic GMP dependant protein kinase (PKG) regulates smooth muscle relaxation. Examination of phosphoproteomes of various smooth muscles identified a distinct subset of early protein targets for PKG. Several were identified in the mouse and human genome, including CHASM, a novel protein containing a previously unidentified motif that is highly conserved in the smoothelin family of smooth muscle specific proteins. When added to permeabilized smooth muscles, CHASM causes calcium desensitization and relaxation in a phosphorylation dependant manner. The degree of sequence divergence of the intervening non-conserved amino acids within the CHASM motif region suggests that CHASM and the smoothelins may be part of a larger family of smooth muscle specific proteins that are important in the mediating the actions of cGMP/PKG. To directly test this hypothesis we have deleted the CHASM gene and obtained CHASM null mice.

平滑肌在多种生理过程中发挥着重要作用，尽管每条平滑肌的基本功能都是相同的，即收缩和放松、机械性能和平滑肌类型之间对激素、神经递质和药物的反应差异很大。决定平滑肌收缩特性的因素包括质膜特性，信号转导蛋白的比例和补充，收缩装置本身的组成。这些成分正常混合的改变被认为是几种物质的分子基础。涉及平滑肌的人类疾病包括高血压、野马痉挛、性功能障碍、胃肠道疾病和青光眼。我们的假设是，通过研究分子过程哪些个体平滑肌通常对刺激做出反应将导致更有选择性的治疗方法这些疾病。最近完成的人类和小鼠基因组结合先进的质谱技术为探测细胞信号转导途径提供了新的机会。在这个提案中，我们将采用蛋白质组学、肌肉生理学、分子生物学、免疫组织化学和小鼠遗传学以确定 cGMP 的分子机制通过激活环 GMP 依赖性蛋白激酶 (PKG) 调节平滑肌松弛。对各种平滑肌磷酸化蛋白质组的检查发现了早期蛋白质的独特子集 PKG 的目标。在小鼠和人类基因组中发现了几种，其中包括 CHASM，一种新型的含有先前未识别的基序的蛋白质，该基序在平滑肌蛋白家族中高度保守肌肉特异性蛋白质。当添加到透化的平滑肌中时，CHASM 会产生钙以磷酸化依赖性方式脱敏和松弛。序列分歧程度 CHASM 基序区域内插入的非保守氨基酸表明 CHASM 和平滑蛋白可能是平滑肌特异性蛋白大家族的一部分，这些蛋白在介导 cGMP/PKG 的作用。为了直接检验这个假设，我们删除了 CHASM 基因并获得 CHASM 无效小鼠。