Dynamic Proteomics of Individual Cancer Cells in Response to a Drug

Why do seemingly identical cells respond differently to a drug? To address this, we studied the dynamics and variability of the protein response of human cancer cells to a chemotherapy drug, camptothecin

A. A. Cohen

2008

Scholarcy highlights

  • We used two rounds of CD tagging with the red fluorescent protein mCherry to obtain a cell clone with red fluorescence in the cytoplasm and stronger red fluorescence in the nucleus
  • We introduced an enhanced yellow fluorescent protein into the red-tagged cells by an additional round of CD tagging, expanded the yellow-tagged cells into clones, and identified the yellow-tagged proteins
  • Dynamic changes as small as 20 to 30% in tagged protein intensity typically can be accurately detected by using the present assay in individual cells
  • This study provided a view of the response to a drug in space and time for about 1000 proteins in individual cells
  • We identified an RNA helicase and a DNA replication factor, whose dynamics varied widely between cells, in a way that corresponds to cell fate: They rise in cells that survive and decrease in cells that die
  • Understanding the human cell as a dynamical system will require viewing it on several levels, including mRNA and protein levels, modifications, and localizations, in individual living cells over time

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