Your browser version may not work well with NCBI's Web applications. More information here...
1: Science. 2007 Aug 3;317(5838):663-6.
Related Articles, Links
Click here to read Click here to read
Forced unfolding of proteins within cells.

Johnson CP, Tang HY, Carag C, Speicher DW, Discher DE.

Biophysical Engineering Lab, University of Pennsylvania, Philadelphia, PA 19104, USA.

To identify cytoskeletal proteins that change conformation or assembly within stressed cells, in situ labeling of sterically shielded cysteines with fluorophores was analyzed by fluorescence imaging, quantitative mass spectrometry, and sequential two-dye labeling. Within red blood cells, shotgun labeling showed that shielded cysteines in the two isoforms of the cytoskeletal protein spectrin were increasingly labeled as a function of shear stress and time, indicative of forced unfolding of specific domains. Within mesenchymal stem cells-as a prototypical adherent cell-nonmuscle myosin IIA and vimentin are just two of the cytoskeletal proteins identified that show differential labeling in tensed versus drug-relaxed cells. Cysteine labeling of proteins within live cells can thus be used to fluorescently map out sites of molecular-scale deformation, and the results also suggest means to colocalize signaling events such as phosphorylation with forced unfolding.

Publication Types:
PMID: 17673662 [PubMed - indexed for MEDLINE]