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. 2013 Dec 8;289(5):2577–2588. doi: 10.1074/jbc.M113.534453

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FIGURE 1. — The UBA2 domain of hHR23A is not sufficient for stable Vpr binding. Analytical gel-filtration profiles of hHR23A (A), UBA2 (hHR23A residues 311–363, B), NusA-Vpr (C), NusA-Vpr mixed with hHR23A (D), and NusA-Vpr mixed with UBA2 (E). Elution volumes are indicated above and fraction numbers below the peaks. F, SDS-PAGE analysis of the fractions indicated in A–E, with lane numbers indicating the fraction numbers. G, SEC-MALS analysis of purified hHR23A (blue) and the hHR23A-Vpr complex (after cleavage of the NusA tag, red). Elution profiles are shown by ○, and the weight-averaged molecular masses obtained from the SEC-MALS measurements are shown by ▵ across the peaks. Inset, SDS-PAGE of free (center lane) and the Vpr-complexed (right lane) hHR23A NMR samples used to obtain the ¹H-¹⁵N HSQC spectra shown in Fig. 2A.