CDy2 stains mitochondria in live muscle cells and target protein was expected to covalently labeled through CA group of CDy2. The fluorescent property of CDy2 allowed to monitor the binding protein from SDS-gel and the biomarker of CDy2 was elucidated as ALDH2.
■ Cell Image
Identification of protein binders in vitro and in living cells. (a) Chemical structure of CDy2 for labeling a target protein in living cells. (b) Myoblasts (MB) or myotubes (MT) were incubated with CDy2 (500 nM) for 30 min and imaged with a fluorescent microscope. Then, cells were lysed for in-gel fluorescence analysis (lex=530 nm, lex=580 nm) (c) Western blot analysis (WB). (d) Two-dimensional gel analysis for the identification of the labeled protein.
■ Related probes: E26, I25, I31
■ Known Property differentiated muscle cell probe ■ Application Immunofluorescence ■ Cell selectivity mechanism POLD (ALDH2 in mitochondria)
■ Reference
1.
Binding of fluorophores to proteins depends on the cellular environment, Kim, Y. K.; Lee, J. S.; Bi, X.; Ha, H. H.; Ng, S. H.; Ahn, Y. H.; Lee, J. J.; Wagner, B. K.; Clemons, P. A.; Chang, Y. T.* Angew. Chem., Int. Ed. Engl. 2011, 50, 2761-2763.
2.
Small-molecule fluorophores to detect cell-state switching in the context of highthroughput screening, Wagner, B. K.; Carrinski, H. A.; Ahn, Y. H.; Kim, Y. K.; Gilbert, T. J.; Fomina, D. A.; Schreiber, S. L.; Chang, Y. T.; Clemons, P. A. J. Am. Chem. Soc. 2008, 130, 4208-4209.
3.
Site-selective labeling at Cys302 of aldehyde dehydrogenase unveils a selective mitochondrial stain, Kim, Y. K.; Lee, J. K.; Lee, J. S.; Yoon, C. N.; Chang, Y. T.* Mol. Biosyst. 2011, 7, 2375-2378.