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. Author manuscript; available in PMC: 2016 Oct 13.
Published in final edited form as: Nature. 2016 Apr 13;532(7599):398–401. doi: 10.1038/nature17433

Figure 4. Kinetic competition between USP14 and the proteasome.

Figure 4

a, Proteasome (1.2 μM) in the presence or absence of USP14 (17 μM) was mixed with Ubn-NCB1 (~0.47 μM) in a quench flow device. (Top) Degradation of NCB1 conjugates is detectable by approximately 15 s. (Bottom) Deubiquitination of Ub4~6-NCB1 can be observed by 100 ms or earlier as Ub2~3-NCB1 species are generated. b, Quantification of Ubn-NCB1 degradation from a. c, Quantification of Ub3- or Ub2-NCB1 species generated by USP14-mediated deubiquitination. USP14-derived reaction products can appear as early as hundreds of ms. Collected samples were analysed by SDS–PAGE/immunoblotting, using antibody to HA. Note that, for b, axes are logarithmic in scale.