INTRINSIC PROBES OF SMOOTH MUSCLE MYOSIN DYNAMICS

平滑肌肌球蛋白动力学的内在探针

基本信息

批准号：
7172332
负责人：
CHRISTOPHER L. BERGER
金额：
$ 35.84万
依托单位：
UNIVERSITY OF VERMONT & ST AGRIC COLLEGE
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-02-01 至 2010-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7172332
关键词：
ATP Hydrolysis ATP phosphohydrolase ATP-G-actin Actins Active Sites Address Affinity Binding Biochemical Biological Clocks Cardiovascular Diseases Catalytic Domain Chemicals Cleaved cell Coupled Cytokinesis DNA Sequence Rearrangement Disease Engineering Event F-Actin Familial Hypertrophic Cardiomyopathy Fluorescence Fluorescent Probes Goals Hydrolysis Inherited Kinetics Lead Link Location Mediating Molecular Molecular Motors Monitor Motion Motor Movement Muscle Muscle Contraction Mutation Myosin ATPase Nucleotides Organelles Point Mutation Positioning Attribute Process Production Protein Isoforms Proteins Relative (related person)Reporting Research Research Personnel Series Signal Transduction Site Skeletal system Smooth Muscle Myosins Solutions Staging Structure Testing Thinking Time Tryptophan Upper arm actin 2 cell motility design desire inorganic phosphate interest programs research study response

项目摘要

DESCRIPTION (provided by applicant): The ultimate goal of this proposal is to examine conformational changes at specific locations within smooth muscle myosin using intrinsic tryptophan fluorescence. Smooth muscle myosin constructs will be genetically engineered to contain either a single tryptophan or a pair of tryptophans at the desired site of interest, which will provide a unique intrinsic fluorescence signal reporting local conformational and structural changes in response to nucleotide binding, ATP hydrolysis, actin-binding, and lever arm movement. These experiments are complementary to the ongoing structural studies in the field, allowing us to explicitly test predictions about domain movements and structural rearrangements during critical steps in the contractile cycle of smooth muscle myosin. Thus we will be able to correlate structural changes in myosin with functional consequences, which relates directly to certain cardiovascular disease. For example, FHC (familial hypertrophic cardiomyopathy) is an inherited, often lethal disease caused by point mutations at key structures within myosin critical to its proper functioning as a molecular motor. We will be examining structural changes in myosin in regions of the molecule directly impacted by mutations that underlie FHC. This will lead to a better understanding of the disease, and thus to better treatment options as well. Therefore, this proposal offers a unique opportunity to critically test fundamental questions about the molecular mechanism of muscle contraction that have not been previously accessible by other spectroscopic probe studies, and the results will have important implications for serious disease states such as FHC.

描述（由申请人提供）：该提案的最终目标是使用内在的色氨酸荧光检查平滑肌肌球蛋白中特定位置的构象变化。平滑肌肌球蛋白构建体将经过基因设计，可以在感兴趣的所需位点包含一对色氨酸或一对色氨酸，这将提供独特的固有荧光信号，报告局部构象和结构变化，以响应核苷酸结合，ATP水解，肌动蛋白结合，肌动蛋白结合以及杠杆武器运动。这些实验与现场正在进行的结构研究相辅相成，使我们能够在平滑肌肌球蛋白的收缩循环中明确测试有关域运动和结构重排的预测。因此，我们将能够将肌球蛋白的结构变化与功能后果相关联，这与某些心血管疾病直接相关。例如，FHC（家族性肥大性心肌病）是一种遗传性的，通常是由肌球蛋白中关键结构的点突变引起的致命疾病，对其作为分子运动的正常功能至关重要。我们将检查直接受FHC突变影响的分子区域中肌球蛋白的结构变化。这将使人们对疾病有更好的了解，从而更好地治疗选择。因此，该提案提供了一个独特的机会，可以严格测试有关肌肉收缩分子机制的基本问题，而其他光谱探针研究以前尚未获得，结果将对诸如FHC等严重疾病状态（例如FHC）具有重要意义。