FLOURESCENT TOOLS FOR THE STUDY OF METALLOTHIONEIN

描述(申请人提供)：我们将为生命科学家提供研究金属硫蛋白和金属硫蛋白-锌信号的荧光工具。近年来，金属硫蛋白-锌信号已成为许多生死信号通路的中心步骤，包括增殖、凋亡和转移。可以说，MT-锌信号在普遍性和重要性上与钙-钙调蛋白、IP3和一氧化氮相媲美。在作用中，金属硫蛋白(MT)可以在氧化或亚硝化刺激下将7个锌离子释放到溶液中(图1)。相反，非金属蛋白，硫蛋白(T)可以隔离7个锌离子，终止锌信号。更耐人寻味的是，T和MT都显示出穿过质膜和细胞器膜的证据，从而允许锌信号在这些隔室之间转置。我们将在特定位置运送标记有荧光记者的T和MT蛋白。这些记者构成荧光共振能量转移(FRET)系统，当锌被结合或释放时，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骗到组织中，然后从循环中进入胞浆，它们可以用于体外和体内研究。有了合适的内窥镜和透皮共聚焦或双光子显微镜，MT/T信号有可能在体内和体外被成像。MT/T和锌信号通路在细胞损伤和死亡途径以及转移和健康细胞生长中都是不可或缺的。我们的工具将促进所有这些领域的研究，并为诊断和治疗医学科学的进步和发展做出贡献。

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.