Lanthanide-Binding Tags for 3D X-ray Imaging of Proteins in Cells at Nanoscale Resolution

被引:37
作者
Victor, Tiffany W. [1 ,2 ]
O'Toole, Katherine H. [3 ]
Easthon, Lindsey M. [3 ]
Ge, Mingyuan [1 ]
Smith, Randy J. [1 ]
Huang, Xiaojing [1 ]
Yan, Hanfei [1 ]
Chu, Yong S. [1 ]
Chen, Si
Gursoy, Doga [4 ,5 ]
Ralle, Martina [6 ]
Imperiali, Barbara [7 ]
Allen, Karen N. [3 ]
Miller, Lisa M. [1 ,2 ]
机构
[1] Brookhaven Natl Lab, Natl Synchrotron Light Source 2, Upton, NY 11973 USA
[2] SUNY Stony Brook, Dept Chem, Stony Brook, NY 11794 USA
[3] Boston Univ, Dept Chem, Boston, MA 02215 USA
[4] Argonne Natl Lab, Adv Photon Source, Lemont, IL 60439 USA
[5] Northwestern Univ, Dept Elect Engn & Comp Sci, Evanston, IL 60298 USA
[6] Oregon Hlth & Sci Univ, Dept Mol & Med Genet, Portland, OR 97239 USA
[7] MIT, Dept Biol, Cambridge, MA 02139 USA
基金
美国能源部; 美国国家卫生研究院;
关键词
PROTEASOME PATHWAY; OMPA; MICROSCOPY; PEPTIDES; DISEASE; ORIGIN;
D O I
10.1021/jacs.9b11571
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We report the application of lanthanide-binding tags (LBTs) for two- and three-dimensional X-ray imaging of individual proteins in cells with a sub-15 nm beam. The method combines encoded LBTs, which are tags of minimal size (ca. 15-20 amino acids) affording high-affinity lanthanide ion binding, and X-ray fluorescence microscopy (XFM). This approach enables visualization of LBT-tagged proteins while simultaneously measuring the elemental distribution in cells at a spatial resolution necessary for visualizing cell membranes and eukaryotic subcellular organelles.
引用
收藏
页码:2145 / 2149
页数:5
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