Lanthanide-Binding Tags for 3D X-ray Imaging of Proteins in Cells at Nanoscale Resolution.
Tiffany W VictorKatherine H O'TooleLindsey M EasthonMingyuan GeRandy J SmithXiaojing HuangHanfei YanYong S ChuSi ChenDoga GursoyMartina RalleBarbara ImperialiKaren N AllenLisa M MillerPublished in: Journal of the American Chemical Society (2020)
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.
Keyphrases
- single molecule
- high resolution
- induced apoptosis
- cell cycle arrest
- energy transfer
- signaling pathway
- computed tomography
- magnetic resonance
- stem cells
- binding protein
- single cell
- living cells
- amino acid
- atomic force microscopy
- cell death
- cell therapy
- transcription factor
- high speed
- fluorescence imaging
- monte carlo
- fluorescent probe