最近,伦敦帝国大学和埃因霍温科技大学的研究人员利用绿色荧光蛋白(green fluorescent proteins,GFPs)开发出一种新的分子传感器——荧光共振能量转移传感器(fluorescence resonance energy transfer based sensor, FRET sensor),首次使科学家能精确测出细胞内的锌元素总浓度和所在位置,从而可以使科学家更好的了解某些疾病,如2型糖尿病等。这项研究发表在8月30日出版的Nature Methods杂志上。
锌元素参与机体许多生理反应,机体细胞中总蛋白量的5%蛋白质都参与了锌元素的运输。在这项研究中,研究人员利用GFPs测出每个细胞中所有锌离子之间的距离以及锌离子浓度.
FRET传感器是由有两种绿色荧光蛋白组成,研究人员使其中一种蛋白质发射一定波长的光,而另一种蛋白则收集前一种蛋白发出的光线。当传感器接触锌离子时,两种蛋白发生分离,而且其发射光线的能力变弱。研究人员利用荧光显微镜可以检测这两种蛋白质发出光线的波长,从而显示出细胞内呈彩色斑块的锌离子的分布区域。
生物谷推荐原文出处:
Nature Methods Published online: 30 August 2009 | doi:10.1038/nmeth.1368
Genetically encoded FRET sensors to monitor intracellular Zn2+ homeostasis
Jan L Vinkenborg1, Tamara J Nicolson2, Elisa A Bellomo2, Melissa S Koay1, Guy A Rutter2 & Maarten Merkx1
1 Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, Eindhoven, The Netherlands.
2 Section of Cell Biology, Division of Medicine, Imperial College London, London, UK.
Top of pageWe developed genetically encoded fluorescence resonance energy transfer (FRET)-based sensors that display a large ratiometric change upon Zn2+ binding, have affinities that span the pico- to nanomolar range and can readily be targeted to subcellular organelles. Using this sensor toolbox we found that cytosolic Zn2+ was buffered at 0.4 nM in pancreatic cells, and we found substantially higher Zn2+ concentrations in insulin-containing secretory vesicles.
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