Molecular and Functional Characterization of Mucolipin-1

Mucolipin-1 的分子和功能表征

• 批准号: 7166098
• 负责人: ABIGAIL A SOYOMBO-SHOOLA
• 金额: $ 13.87万
• 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/7166098
• 关键词: Appendix; Arts; Binding; Biochemistry; Biomedical Research; C-terminal; Cathepsins B; Cations; Cell Fractionation; Cell membrane; Cell physiology; Cells; Cleaved cell; Clinical; Complementary DNA; Complex; Confocal Microscopy; Core Facility; Defect; Density Gradient Centrifugation; Development; 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; experience; improved; inhibitor/antagonist; late endosome; lipid metabolism; member; prevent; protease inhibitor E64d; protein function; receptor; research study; trafficking

DESCRIPTION (provided by applicant): 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-art core 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 an 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 the following hypothesis: 1) Characterize the molecular determinants of lysosomal targeting and proteolytic cleavage, 2) identify which lipases are activated by ML1, and 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.

描述（由申请人提供）：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一个独立的研究者在神经退行性疾病和钙（2+）信号研究。申请人对生物医学研究事业表现出坚定的承诺，并在细胞和分子生物学、生物化学以及溶酶体贮积病的基因靶向方面获得了广泛的既往培训和专业知识，但是，她认为，在本拟议研究的背景下，在概述的成像和电生理技术方面接受一段时间的额外培训将大大提高她作为独立科学家的竞争力。申请人的环境非常适合支持她的持续职业发展，可以使用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+）信号传导和离子通道功能调节方面的专家，在指导年轻研究人员方面拥有丰富的经验。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粘脂沉积症IV型（MLIV），一种脂质储存神经退行性疾病。MLIV在细胞水平上定义为脂质的溶酶体蓄积和晚期内体/杂合细胞器形成成熟溶酶体的阻滞。ML 1的确切功能尚不清楚。在初步研究中，我们已经表明，ML 1是一个H+选择性阳离子通道，在溶酶体中特异性裂解。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活性的作用。我们现在假设ML 1是一种双功能蛋白，通过介导溶酶体pH值的改变来调节溶酶体脂质代谢。在没有功能性ML 1的情况下，脂质代谢异常。以下特定目的将检验以下假设：1）表征溶酶体靶向和蛋白水解裂解的分子决定簇，2）鉴定哪些脂肪酶被ML 1激活，3）表征ML 1与MLS以及与可能在复合物中起作用的其他蛋白质的相互作用。这些研究的结果将为这种疾病的细胞和治疗提供基础。