Reyes-Turcu, F. E., Shanks, J. R., Komander, D., Wilkinson, K. D. (2008) Recognition of polyubiquitin isoforms by the multiple ubiquitin binding modules of isopeptidase T. JOURNAL OF BIOLOGICAL CHEMISTRY, 283 (28). pp. 19581-19592. ISSN 0021-9258

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Abstract

The conjugation of polyubiquitin to target proteins acts as a signal that regulates target stability, localization, and function. Several ubiquitin binding domains have been described, and while much is known about ubiquitin binding to the isolated domains, little is known with regard to how the domains interact with polyubiquitin in the context of full-length proteins. Isopeptidase T(IsoT/USP5) is a deubiquitinating enzyme that is largely responsible for the disassembly of unanchored polyubiquitin in the cell. IsoT has four ubiquitin binding domains: a zinc finger domain (ZnF UBP), which binds the proximal ubiquitin, a UBP domain that forms the active site, and two ubiquitin-associated (UBA) domains whose roles are unknown. Here, we show that the UBA domains are involved in binding two different polyubiquitin isoforms, linear and K48-linked. Using isothermal titration calorimetry, we show that IsoT has at least four ubiquitin binding sites for both polyubiquitin isoforms. The thermodynamics of the interactions reveal that the binding is enthalpy-driven. Mutation of the UBA domains suggests that UBA1 and UBA2 domains of IsoT interact with the third and fourth ubiquitins in both polyubiquitin isoforms, respectively. These data suggest that recognition of the polyubiquitin isoforms by IsoT involves considerable conformational mobility in the polyubiquitin ligand, in the enzyme, or in both.

Item Type:	Article
Authors (ICR Faculty only):	Komander, David
All Authors:	Reyes-Turcu, F. E., Shanks, J. R., Komander, D., Wilkinson, K. D.
Uncontrolled Keywords:	UBA DOMAIN; DEUBIQUITINATING ENZYME; STRUCTURAL BASIS; CRYSTAL-STRUCTURE; PROTEIN; CHAINS; COMPLEX; FAMILY; TETRAUBIQUITIN; MONOUBIQUITIN
Research teams:	Closed research groups > Other closed groups
Depositing User:	Users 10 not found.
Date Deposited:	21 Jul 2008 10:39
Last Modified:	19 Aug 2011 09:23
URI:	http://publications.icr.ac.uk/id/eprint/6636

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