(A) Gel filtration elution profiles of Naa50 with NatA in S. pombe (top),human (middle), and cross-species between yeast S. pombe and S. cerevisiae (bottom), using a Superdex S200 column, with either 200 mM or 1M NaCl in sizing buffer. Coomassie-stained SDS–PAGE of peak fractions are shown to the right of the chromatograms.
(B) Fluorescence polarization assays with NatA titrated into fluorescein-5-maleimide labeled Naa50 in both S. pombe (left) and human (right) systems. The data is fit to calculate a dissociation constant between NatA and Naa50.
(C) Differential scanning fluorimetry assays of NatA alone or with Naa50 in both yeast and human systems. Recorded melting temperature transitions are indicated.