Abstract
Bioorthogonal chemical proteomics is a valuable method to identify enzyme-specific substrates, a challenging task by traditional biochemical standards. The addition of recombinant enzyme and alkynyl chemical reporter to complex protein mixtures, such as cell lysates, allows the detection and identification of modified substrates. Proteins that have been modified with the chemical reporter can be selectively labeled with fluorescent dyes for detection or affinity tags for biochemical enrichment and subsequent identification by mass spectrometry. Here, we describe the detection and identification of substrates of the lysine acetyltransferase p300 in nuclear extracts using the chemical reporter 4-pentynoyl-CoA.
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Grammel, M., Hang, H.C. (2013). Identification of Lysine Acetyltransferase Substrates Using Bioorthogonal Chemical Proteomics. In: Hake, S., Janzen, C. (eds) Protein Acetylation. Methods in Molecular Biology, vol 981. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-305-3_16
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DOI: https://doi.org/10.1007/978-1-62703-305-3_16
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-304-6
Online ISBN: 978-1-62703-305-3
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