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. 2016 Nov 4;26(11):1197–1211. doi: 10.1038/cr.2016.123

Figure 2.

Figure 2

Sre1-CTD exists as a dimer of dimer. (A) The Sre1-CTD may exist as an oligomer. The Sre1-CTD (residues 628-896) was eluted from SEC at a position corresponding to the molecular weight of a tetramer. The cross-linked Sre1-CTD, which showed a molecular weight of ∼120 kDa on SDS-PAGE, was eluted at a similar position to the WT protein on SEC. (B) Static light scattering (SLS) analysis of the Sre1-CTD. SLS analysis of the WT and cross-linked samples suggested that the Sre1-CTD may form a less stable tetramer as the cross-linked protein was measured with a molecular weight of a tetramer, whereas the WT protein exhibited a slightly reduced molecular weight. (C) Analytical ultracentrifugation analysis (AUC) of the Sre1-CTD. Consistent with the SLS analysis, the sedimentation velocity AUC (AUC-SV) analysis of the WT and cross-linked Sre1-CTD supported the formation of an unstable tetramer of Sre1-CTD. (D) Sedimentation equilibrium AUC analysis of the Sre1-CTD. The AUC-SE analysis yielded a dissociation coefficient (Kd) of 1 μM between tetramer and dimer, suggesting a dimer of dimer organization of the Sre1-CTD. See also Supplementary information, Figure S3.