@article{f7d2d80dcc55415086eeca0fc3742a7c,
title = "The differential modulation of USP activity by internal regulatory domains, interactors and eight ubiquitin chain types",
abstract = "Ubiquitin-specific proteases (USPs) are papain-like isopeptidases with variable inter- and intramolecular regulatory domains. To understand the effect of these domains on USP activity, we have analyzed the enzyme kinetics of 12 USPs in the presence and absence of modulators using synthetic reagents. This revealed variations of several orders of magnitude in both the catalytic turnover (k cat) and ubiquitin (Ub) binding (K M) between USPs. Further activity modulation by intramolecular domains affects both the k cat and K M, whereas the intermolecular activators UAF1 and GMPS mainly increase the k cat. Also, we provide the first comprehensive analysis comparing Ub chain preference. USPs can hydrolyze all linkages and show modest Ub-chain preferences, although some show a lack of activity toward linear di-Ub. This comprehensive kinetic analysis highlights the variability within the USP family.",
author = "Faesen, \{Alex C.\} and Luna-Vargas, \{Mark P.A.\} and Geurink, \{Paul P.\} and Marcello Clerici and Remco Merkx and \{Van Dijk\}, \{Willem J.\} and Hameed, \{Dharjath S.\} and \{El Oualid\}, Farid and Huib Ovaa and Sixma, \{Titia K.\}",
note = "Funding Information: We thank Martin A. Cohn and Alan D'Andrea for USP1, USP12, USP46, and UAF1 ccDNA, Annette Dirac and Rene Bernards for USP4 and USP8 cDNA, Elisabetta Citterio for USP21 and USP39 cDNA, Carlos Lopex-Otin for USP30 cDNA, and Erik Meulmeester and Frauke Melchior for USP25 cDNA. We thank Ovaa and Sixma group members for discussion, sharing reagents, and critical reading of the manuscript. This work was supported by grants from the Dutch Cancer Society, the Netherlands Organization for Scientific Research (VIDI grant), and EU-Rubicon and NWO-CW ECHO 700.59.009, KWF-2008-4014, and STW. USP expression and purification were performed by M.P.A.L.V. with assistance from W.J.v.D; enzyme assays were designed and analyzed by A.C.F. and executed by M.P.A.L.V and A.C.F.; USP1, USP12, USP46, and UAF1 were expressed, purified, and analyzed by M.C.; UbAMC was designed and synthesized by R.M.; di-Ubs were designed by F.E. and H.O. and synthesized by D.S.H.; FP reagents were designed by F.E., P.P.G., and H.O. and synthesized by P.P.G. and F.E.; FP reagent assays were performed by P.P.G.; H.O. supervised all synthesis efforts and FP experiments; T.K.S. designed and supervised the USP project; data analysis and manuscript writing were by A.C.F. with M.P.A.L.V. and T.K.S. ",
year = "2011",
month = dec,
day = "23",
doi = "10.1016/j.chembiol.2011.10.017",
language = "English",
volume = "18",
pages = "1550--1561",
journal = "Chemistry and Biology",
issn = "1074-5521",
publisher = "Elsevier Inc.",
number = "12",
}