Single Fluorescent Protein-Based Indicators for Zinc Ion (Zn(2+)).
Single Fluorescent Protein-Based Indicators for Zinc Ion (Zn(2+)).
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DOI：
10.1021/acs.analchem.6b01653
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发表时间：
2016-09-20
影响因子：
7.4
通讯作者：
Ai HW
中科院分区：
文献类型：
--
作者：
Chen Z;Ai HW
基因编码的Zn 2+荧光指示剂（GEZI）是研究哺乳动物细胞中Zn 2+稳态和信号传导的非常有吸引力的研究工具。目前大多数GEZI是基于与Zn 2+结合基序融合的一对荧光蛋白（FP）之间的Förster共振能量转移（FRET）。这种基于FRET的GEZI的一个缺点是它们的宽光谱轮廓带宽，在监测多个目标或参数时产生挑战。为了解决这个问题，我们设计了一组基于单个蓝绿色和红色FP的增强型GEZI，可用于监测胰腺INS-1 E β细胞中的亚细胞Zn 2+扩散和葡萄糖诱导的Zn 2+分泌。这些GEZI提供了简单的强度测量，兼容性，大的动态范围，并相对较小的分子尺寸，使其成为有价值的补充分子工具箱成像Zn 2+。
Genetically encoded fluorescent Zn2+ indicators (GEZIs) are highly attractive research tools for studying Zn2+ homeostasis and signaling in mammalian cells. Most current GEZIs are based on Förster resonance energy transfer (FRET) between a select pair of fluorescent proteins (FPs) fused with Zn2+-binding motifs. One drawback of such FRET-based GEZIs is their broad spectral profile bandwidths, creating challenges when monitoring multiple targets or parameters. To address this issue, we have engineered a group of intensiometric GEZIs based on single teal and red FPs that can be utilized to monitor subcellular Zn2+ diffusion and glucose-induced Zn2+ secretion in pancreatic INS-1E β-cells. These GEZIs offer the simplicity of intensiometric measurements, compatibility in multicolor imaging, large dynamic ranges, and relatively small molecular sizes, making them valuable additions to the molecular toolbox for imaging Zn2+.