DNA Binding in High Salt: Analysing the Salt Dependence of Replication Protein A3 from the Halophile Haloferax volcanii
Figure 5
(a) Size exclusion profiles of HvRPA3-NTD in the presence (dashed line) and absence (solid line) of equimolar 18mer ssDNA in 0.2, 1, and 3 M KCl. The elution profile of the oligonucleotide alone is shown (dotted line). The axis shows elution volume (mls) and the axis absorbance at 280 nm. For clarity the peaks containing protein and protein/oligonucleotide have been scaled relative to the oligonucleotide alone. Peak heights (mAu) were 0.2 M 6.2/6.7, 1 M 7.4/40.5, and 3 M 10.1/59.3 (NTD alone/NTD+18mer ssDNA). Arrows indicate the presence of ssDNA in complex (dotted line) and alone (dashed line). (b) SDS-PAGE analysis of the indicated fractions in the presence and absence of oligonucleotide. Increased salt concentrations induce a degree of warping in the gels.
