Abstract
Proteins targeted for degradation by the mycobacterial proteasome are covalently modified with prokaryotic ubiquitin-like protein (Pup) in a process termed “pupylation.” Despite its name, Pup is only ubiquitin-like in function and not sequence or structure. Furthermore, the enzymology of pupylation appears to be distinct from protein modification by ubiquitin (Ub) and other ubiquitin-like proteins (Ubls). Nonetheless, we have adapted methods established in the Ub field for the production of reagents to isolate, identify, and analyze pupylated proteins in mycobacteria. These methods can be modified to study specific pupylated proteins in various Pup-bearing bacteria or to identify posttranslational modifiers in other prokaryotes.
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Acknowledgments
We thank Andrew Darwin and Tony Huang for critical review of this chapter. This work was supported by the NIH (1R01HL092774) and the Irma T. Hirschl Charitable Trust. K. Heran Darwin, Ph.D. holds an Investigators in the Pathogenesis of Infectious Disease Award from the Burroughs Wellcome Fund.
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Burns, K.E., Darwin, K.H. (2012). Pupylation: Proteasomal Targeting by a Protein Modifier in Bacteria. In: Dohmen, R., Scheffner, M. (eds) Ubiquitin Family Modifiers and the Proteasome. Methods in Molecular Biology, vol 832. Humana Press. https://doi.org/10.1007/978-1-61779-474-2_10
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DOI: https://doi.org/10.1007/978-1-61779-474-2_10
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