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 study what features in INO80GDEU are required to achieve UCH-L5 inhibition, we analyzed its differences with the activating DEUBAD domain of RPN13. The most striking difference between the DEUBAD domains of RPN13 and INO80G is the C-terminal part where three helices α6–α8 in RPN13DEU change to the extended α6 in INO80GDEU that packs against the CD (Figures 5A and S1D). Therefore, we created a shorter version of INO80GDEU, where helix α6 is removed (Figure 1A, residues 39–118, INO80GDEUΔα6). Surprisingly, the truncated protein INO80GDEUΔα6 still inhibited UCH-L5 (Figure 5B), demonstrating that helix α6 is not required for inhibition under these conditions.
Affiliation: Division of Biochemistry and Cancer Genomics Center, The Netherlands Cancer Institute, Plesmanlaan 121, 1066CX Amsterdam, the Netherlands.