Sequential Activation and Deactivation of Protein Function Using Spectrally Differentiated Caged Phosphoamino Acids
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Imperiali_Sequential activation.pdf
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Author(s)
Goguen, Brenda N.
Aemissegger, Andreas
Imperiali, Barbara
Date Issued
June 2011
Journal
Journal of the American Chemical Society
Publisher
American Chemical Society (ACS)
Citation
Goguen, Brenda N., Andreas Aemissegger, and Barbara Imperiali. “Sequential Activation and Deactivation of Protein Function Using Spectrally Differentiated Caged Phosphoamino Acids.” Journal of the American Chemical Society 133.29 (2011): 11038–11041.
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Author's final manuscript
Abstract
Photolabile caging groups, including the 1-(2-nitrophenyl)ethyl (NPE) group, have been applied to probe many biological processes, including protein phosphorylation. Although studies with NPE-caged phosphoamino acids have provided valuable information, these investigations have been limited to the use of only one caged species in a single experiment. To expand the scope of these tools, we have developed an approach for sequentially uncaging two different phosphopeptides in one system, enabling interrogation of multiple phosphorylation events. We present the synthesis of [7-(diethylamino)coumarin-4-yl]methyl (DEACM)-caged phosphorylated serine, threonine, and tyrosine building blocks for Fmoc-based solid-phase peptide synthesis to allow convenient incorporation of these residues into peptides and proteins. Exposure of DEACM- and NPE-caged phosphopeptides to 420 nm light selectively releases the DEACM group without affecting the NPE-caged peptide. This then enables a subsequent irradiation event at 365 nm to remove the NPE group and liberate a second phosphopeptide. We demonstrate the versatility of this general sequential uncaging approach by applying it to control Wip1 phosphatase with two wavelengths of light.
MIT Department
Massachusetts Institute of Technology. Department of Biology
Massachusetts Institute of Technology. Department of Chemistry
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DOI of Published Version
https://doi.org/10.1021/ja2028074
