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. 2016 Dec;196(3):437–447. doi: 10.1016/j.jsb.2016.09.011

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© 2016 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

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Fig. 5 — The two step UAF1 binding is conserved in USP1. A) SEC-MALLS analysis of USP1/UAF1_FL (6 μM) shows a molecular weight corresponding to the 1:2 complex. B) Qualitative SPR analysis shows that USP1 has conserved the two-step binding to UAF1_FL, with similar kinetics. C) Fitting of quantitative SPR analysis of UAF1_FL to USP1 obtained from 15 successive injections of UAF1_FL to quantify the K_D values. Curve is fit to a two-step binding model with affinities as indicated. D) UAF1 binds USP12 at the fingers sub-domain which activates USP12 and then UAF1 also binds on the backside of the ubiquitin binding cleft.