Author: Swatek, Kirby N.; Aumayr, Martina; Pruneda, Jonathan N.; Visser, Linda J.; Berryman, Stephen; Kueck, Anja F.; Geurink, Paul P.; Ovaa, Huib; van Kuppeveld, Frank J. M.; Tuthill, Tobias J.; Skern, Tim; Komander, David
Title: Irreversible inactivation of ISG15 by a viral leader protease enables alternative infection detection strategies Document date: 2018_3_6
ID: 3s86w4iw_10
Snippet: The cleavage site of Lb pro contrasts all known deISGylases, which cleave ISG15 from substrates, such that both proteins are recycled to the same state as they were before modification (18, 19) . In hindsight, these findings explain the inability of Lb pro to react with the ISG15 suicide probe (in which the reactive warhead would extend beyond the active site) or generate a signal in the ISG15-AMC assay. AMC only fluoresces in the unmodified form.....
Document: The cleavage site of Lb pro contrasts all known deISGylases, which cleave ISG15 from substrates, such that both proteins are recycled to the same state as they were before modification (18, 19) . In hindsight, these findings explain the inability of Lb pro to react with the ISG15 suicide probe (in which the reactive warhead would extend beyond the active site) or generate a signal in the ISG15-AMC assay. AMC only fluoresces in the unmodified form, whereas Lb pro cleavage produces a nonfluorescent GlyGlymodified AMC reagent ( Fig. S2A ).
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