Abstract
Benzophenone photoprobes represent powerful tools for chemical proteomics. Upon UV irradiation, a benzophenone photoprobe can selectively form a covalent bond with its target protein in complex protein mixtures. Thus, photoprobes can be used to profile a wide variety of proteins in complex proteomes. This chapter describes simple protocols to derivatize fluorenylmethyloxycarbonyl (Fmoc)-protected peptide-nucleic-acid adenine (PNA adenine) into a benzophenone photoprobe and its application in photolabeling its target proteins. The method as described does not require specialized equipment for probe synthesis and photolabeling. In addition, the strategy is applicable to recognition motifs other than PNA adenine, such as peptides, to profile their target proteins in complex proteomes.
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Acknowledgments
The authors would like to thank Dr. Skagit Hindi, who was involved in the initial development of our PNA-adenine photoprobes. This work was supported by PSC CUNY grant (PSCREG-37-813). RR-03037 from NCRR/NIH, which supports the research infrastructure at Hunter College, is also acknowledged. We also thank Mr. Kwang Won Cha, Mr. Laurence James, and Ms. Olga Aminova for technical assistance.
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Kawamura, A., Mihai, D.M. (2012). Probing Proteomes with Benzophenone Photoprobes. In: Drewes, G., Bantscheff, M. (eds) Chemical Proteomics. Methods in Molecular Biology, vol 803. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-61779-364-6_6
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DOI: https://doi.org/10.1007/978-1-61779-364-6_6
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