UBP43, an ISG15-specific deconjugating enzyme: expression, purification, and enzymatic assays.
UBP43 is the only deconjugating enzyme for the protein ISGylation system thus far identified. UBP43 activity is not critical for precursor processing, as UBP43-deficent cells can generate ISG15 conjugates upon type I interferon treatment. However, UBP43 deficiency caused a defect in the negative regulation of type I interferon signaling, resulting in enhanced and prolonged activation of Jak/Stat upon type I interferon treatment. This chapter describes the expression, purification, and enzymatic assays for UBP43.
Kim
Keun Il
KI
Department Of Molecular And Experimental Medicine (MEM-L51), The Scripps Research Institute, La Jolla, CA 92037, USA.
Zhang
Dong-Er
DE
D000818:Animals; D003001:Cloning, Molecular; D016207:Cytokines / Q000235:genetics / Q000378:metabolism*; D010450:Endopeptidases / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D006801:Humans; D051379:Mice; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D043222:Ubiquitin Thiolesterase; D014452:Ubiquitins / Q000235:genetics / Q000378:metabolism; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D016207:Cytokines; D011993:Recombinant Fusion Proteins; D014452:Ubiquitins; C074034:ISG15 protein, human; D001616:beta-Galactosidase; D010450:Endopeptidases; C118306:Usp18 protein, mouse; C485560:USP18 protein, human; D043222:Ubiquitin Thiolesterase