FLOURESCENT TOOLS FOR THE STUDY OF METALLOTHIONEIN

用于研究金属硫蛋白的荧光工具

• 批准号: 7111537
• 负责人: CHRISTOPHER J. FREDERICKSON
• 金额: $ 33.85万
• 依托单位: NEUROBIOTEX, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-21 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7111537
• 关键词: metallothionein; proteins; zinc

DESCRIPTION (provided by applicant): We will deliver to life scientists fluorescent tools for the study of metallothionein and metallothionein- zinc signaling. Metallothionein-zinc signals have recently emerged as central step in many life-or-death signal pathways, including proliferation, apoptosis and metastasis. Arguably, MT-zinc signaling rivals calcium-calmodulin, IP3, and nitric oxide in ubiquity and essentiality. In action, the metallated protein, metallothionein (MT) can release 7 zinc ions into solution under oxidative or nitrosative stimulation (Fig 1). Conversely, The non-metalated protein, thionein (T) can sequester 7 zinc ions, terminating a zinc signal. Even more intriguing, both T and MT show evidence of moving through both plasma and organelle membranes, thus allowing zinc signals to be transposed among and between those compartments. We will deliver T and MT proteins that are labeled with fluorescent reporters at specific positions. These reporters constitute fluorescence-resonance-energy-transfer (FRET) systems that change color (emission spectra) when zinc is bound or released (Fig. 2). Because these proteins can be coaxed into tissues and thence into cytosol from the circulation, they can be used for both in vitro and in vivo research. With appropriate endoscopic and transdermal confocal or two-photon microscopy MT/T signaling can potentially be imaged in vivo and in vitro. MT/T and zinc signaling are integral to cell injury and death pathways as well as both metastatic and .healthy cell growth. Our tools will facilitate research in all of those areas and contribute to the advancement and development of diagnostic and therapeutic medical science.

描述（由申请人提供）：我们将向生命科学家提供用于研究金属硫蛋白和金属硫蛋白-锌信号传导的荧光工具。金属硫蛋白-锌信号是细胞增殖、凋亡和转移等多种生死信号通路的关键环节。可以说，MT-锌信号转导在普遍性和重要性方面与钙-钙调蛋白、IP 3和一氧化氮相媲美。在作用中，金属化的蛋白质金属硫蛋白（MT）在氧化或亚硝化刺激下可以释放7个锌离子到溶液中（图1）。相反，非金属化蛋白硫蛋白（T）可以螯合7个锌离子，终止锌信号。更有趣的是，T和MT都显示出穿过血浆和细胞器膜的证据，从而允许锌信号在这些隔室之间和之间转移。我们将提供在特定位置用荧光报告分子标记的T和MT蛋白。这些报告分子构成荧光共振能量转移（FRET）系统，当锌结合或释放时，该系统会改变颜色（发射光谱）（图2）。因为这些蛋白质可以被诱导进入组织，然后从循环进入细胞溶质，所以它们可以用于体外和体内研究。通过适当的内窥镜和经皮共聚焦或双光子显微镜，MT/T信号可能在体内和体外成像。MT/T和锌信号传导是细胞损伤和死亡途径以及转移性和健康细胞生长的组成部分。我们的工具将促进所有这些领域的研究，并有助于诊断和治疗医学科学的进步和发展。