Mechanism of UCH-L5 activation and inhibition by DEUBAD domains in RPN13 and INO80G.
Bottom Line: In this process, large conformational changes create small but highly specific interfaces that mediate activity modulation of UCH-L5 by altering the affinity for substrates.Our results establish how related domains can exploit enzyme conformational plasticity to allosterically regulate DUB activity.These allosteric sites may present novel insights for pharmaceutical intervention in DUB activity.
Affiliation: Division of Biochemistry and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, the Netherlands.Show MeSH
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Mentions: To investigate how RPN13DEU promotes enhanced substrate binding by UCH-L5, we tested the effect of mutations on activity. We first focused on the effect on activation of the ubiquitin-binding residues in UCH-L5. Mutations in these residues lowered the activity substantially, irrespective of the presence of RPN13DEU, indicating that they are primarily important for basic DUB function (Figure 3A). We then tested mutations of RPN13DEU located in the interface with ubiquitin, and found that these provided only a limited contribution to UCH-L5 activation (Figures 2D and S2C).
Affiliation: Division of Biochemistry and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, the Netherlands.