Fig. 1: Characterization of CtNatB and its subunits.
From: Structural basis of Naa20 activity towards a canonical NatB substrate
a–c SEC-MALS analysis of CtNaa20 (a), CtNaa25 (b), and CtNatB (c). The experimentally determined molecular weight of CtNatB, CtNaa25, and CtNaa20 is 136.5 kDa (theoretical molecular is 136.1 kDa), 110.7 kDa (theoretical Mw is 115.1 kDa), and 23.6 kDa (theoretical Mw is 23.1 kDa), respectively. The UV-signals (black) of the corresponding SEC chromatograms are shown together with the light scattering signals (gray) and the mass distributions (black dots). d Isothermal titration calorimetry measurement of CtNatB complex formation. CtNaa20 was titrated by CtNaa25. The signal of one representative measurement is given in differential points (DP) and the dissociation constant Kd and binding enthalpy are given in the table. The heats of dilution of the buffer to CtNaa20 (triangle) and CtNaa25 to buffer (cross) control runs are represented. Measurements were performed in triplicate and the values represent the means and standard deviations. e Electrophoretic mobility shift assay with the tip of CtES27 (expansion segment 27) RNA, CtNaa20, and CtNatB. CtNaa20 and CtNatB were mixed in different ratios with the RNA and free or bound RNA is indicated.
