Molecular and Functional Characterization of Mucolipin-1

Mucolipin-1 的分子和功能表征

• 批准号: 7591030
• 负责人: ABIGAIL A SOYOMBO-SHOOLA
• 金额: $ 14.38万
• 依托单位: UT SOUTHWESTERN MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-05-01 至 2011-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7591030
• 关键词: Binding; Biochemistry; Biomedical Research; C-terminal; Cathepsins B; Cations; Cell Fractionation; Cell membrane; Cell physiology; Cells; Cleaved cell; Clinical; Complementary DNA; Complex; Confocal Microscopy; Defect; Density Gradient Centrifugation; Disease; Environment; Enzymes; Family; Fibroblasts; Ganglioside Sialidase Deficiency Disease; Gene Targeting; Genes; Goals; Hybrids; Image; Individual; Inherited; Insecta; Ion Channel; Ion Channel Protein; K-Series Research Career Programs; Lead; Length; Lipase; Lipids; Lysosomal Storage Diseases; Lysosomes; Maintenance; Mediating; Membrane; Mentors; Metabolic; Molecular; Molecular and Cellular Biology; Mutation; N-terminal; Nerve Degeneration; Neuraxis; Neurodegenerative Disorders; Neuroglia; Neurons; Null Lymphocytes; Organ; Organelles; Palmitoyl Coenzyme A; Palmitoyl-CoA Hydrolase; Pathway interactions; Patients; Percoll; Physiological Processes; Pilot Projects; Process; Proteins; Proteolysis; Regulation; Reporter; Research; Research Personnel; Role; Scientist; Signal Transduction; Site; Sorting - Cell Movement; Staging; Techniques; Testing; Therapeutic; Tissues; Training; Transfection; Tyrosine; Visual; Water; Western Blotting; base; career; career development; experience; improved; inhibitor/antagonist; late endosome; lipid metabolism; member; prevent; protease inhibitor E64d; protein function; receptor; research study; trafficking

The primary goal of this Mentored Career Development Award is to prepare the applicant for a career as an independent investigator in neurodegenerative and Ca(2+) signaling research. The applicant has shown a firm commitment to a biomedical research career and has acquired extensive prior training and expertise in cell and molecular biology, biochemistry, and gene targeting of lysosomal storage diseases, however, she believes that a period of additional training in the outlined imaging and electrophysiological techniques in the context of this proposed study will dramatically improve her competitiveness as an independent scientist. The applicant's environment is ideally suited to support her continued career development, with access to state-of-the-artcore facilities and on-site scientific experts. Her mentor, Dr. Muallem, is an expert in Ca(2+) signaling and in the regulation of ion channel functions and has had extensive experience in mentoring young investigators. The candidate's immediate goal is to elucidate the cellular functions of TRP-ML1 (ML1), and to determine how defects in this lysosomal membrane ion channel lead to mucolipidosis type IV (MLIV), a lipid storage neurodegenerative disorder. MLIV is defined at the cellular level by a lysosomal accumulation of lipids and by a block in the formation of mature lysosomes from late endosomes/hybrid organelles. The exact function of ML1 is unknown. In preliminary studies, we have shown that ML1 is a H+ selective cation channel that is specifically cleaved in the lysosomes. The cleaved form is predominantly seen in native cells and tissues and may function to regulate lipase activity. We now hypothesize that ML1 is a dual function protein that regulates lysosomal lipid metabolism by mediating alterations in lysosomal pH. In the absence of a functional ML1, lipid metabolism is aberrant. The following specific aims will test this hypothesis: 1) Characterize the molecular determinants of lysosomal targeting and proteolytic cleavage, 2) Identify which lipases are activated by ML1, 3) Characterize the interactions of ML1 with MLS and with other proteins that may function in a complex. Results from these studies will provide the cellular and therapeutic basis of this disease.

这个指导职业发展奖的主要目标是准备申请人的职业生涯 作为神经退行性疾病和Ca（2+）信号研究的独立研究者。申请人 表现出对生物医学研究事业的坚定承诺，并获得了广泛的优先 细胞和分子生物学、生物化学和溶酶体基因靶向方面的培训和专业知识 然而，她认为，在概述的成像中进行一段时间的额外训练， 在这项拟议研究的背景下，电生理技术将大大改善她的健康状况。 作为一名独立的科学家。申请人的环境非常适合支持 她的持续职业发展，与访问国家的最先进的核心设施和现场科学 专家她的导师Muallem博士是Ca（2+）信号和离子通道调节方面的专家 他在指导年轻调查员方面具有丰富经验。 候选人的直接目标是阐明TRP-ML 1（ML 1）的细胞功能，并 确定这种溶酶体膜离子通道的缺陷如何导致IV型粘脂沉积症（MLIV）， 一种脂质沉积神经退行性疾病MLIV在细胞水平上由溶酶体膜定义。 脂质的积累，并通过阻止成熟溶酶体的形成， 核内体/杂交细胞器。ML 1的确切功能尚不清楚。在初步研究中， 显示ML 1是在溶酶体中特异性裂解的H+选择性阳离子通道。的 裂解形式主要见于天然细胞和组织中，并可起到调节脂肪酶的作用 活动我们现在假设ML 1是一种调节溶酶体脂质的双重功能蛋白， 在没有功能性ML 1的情况下，脂质代谢通过介导溶酶体pH的改变而发生变化。 新陈代谢异常以下具体目标将检验这一假设：1）描述 溶酶体靶向和蛋白水解裂解的分子决定簇，2）鉴定哪些脂肪酶是 3）表征ML 1与MLS以及与其他蛋白质的相互作用， 在一个复杂的功能。这些研究的结果将提供细胞和治疗基础， 疾病