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Science 3 March 2006:
Vol. 311. no. 5765, pp. 1293 - 1297
DOI: 10.1126/science.1122224
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Reports

Chemical Rescue of a Mutant Enzyme in Living Cells

Yingfeng Qiao,1 Henrik Molina,2 Akhilesh Pandey,2 Jin Zhang,1 Philip A. Cole1*

The restoration of catalytic activity to mutant enzymes by small molecules is well established for in vitro systems. Here, we show that the protein tyrosine kinase Src arginine-388->alanine (R388A) mutant can be rescued in live cells with the use of the small molecule imidazole. Cellular rescue of a viral Src homolog was rapid and reversible and conferred predicted oncogenic properties. Using chemical rescue in combination with mass spectrometry, we confirmed six known Src kinase substrates and identified several new protein targets. Chemical rescue data suggest that cellular Src is active under basal conditions. Rescue of R388A cellular Src provided insights into the mitogen-activated protein kinase pathway. This chemical rescue approach will likely have many applications in cell signaling.

1 Department of Pharmacology and Molecular Sciences, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
2 Department of Biological Chemistry and the Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.

* To whom correspondence should be addressed. E-mail: pcole{at}jhmi.edu

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Science. ISSN 0036-8075 (print), 1095-9203 (online)