Burns Kristin E, Darwin K Heran
Institute for Genetics, University of Cologne, Cologne, Germany.
Methods Mol Biol. 2012;832:151-60. doi: 10.1007/978-1-61779-474-2_10.
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.
被分枝杆菌蛋白酶体靶向降解的蛋白质在一个称为“Pupylation”的过程中与原核泛素样蛋白（Pup）发生共价修饰。尽管其名称如此，但Pup仅在功能上与泛素相似，在序列或结构上并非如此。此外，Pupylation的酶学似乎与泛素（Ub）和其他泛素样蛋白（Ubls）对蛋白质的修饰不同。尽管如此，我们还是采用了在泛素领域建立的方法来生产试剂，以分离、鉴定和分析分枝杆菌中的Pupylated蛋白。这些方法可以进行修改，以研究各种携带Pup的细菌中的特定Pupylated蛋白，或鉴定其他原核生物中的翻译后修饰因子。
