The SOCS-Box of HIV-1 Vif Interacts with ElonginBC by Induced-Folding to Recruit Its Cul5-Containing Ubiquitin Ligase Complex (original) (raw)
Figure 2
Biochemical analysis of the Vif-EloBC interaction.
(A) Schematic representation of the Vif SOCS-box constructs and mutants used in this study. The amino acid sequence is indicated on top, with the mutated residues in gray. (B) Gel filtration binding assay. The Vif SOCS-box and mutated variants were mixed with equimolar amounts of EloBC and run through a gel filtration column, with the UV280 trace shown on top. The eluted fractions were collected and run on a 16% polyacrylamide gel and stained with Coomassie blue (bottom). (C) ITC binding assay. The Vif fusion proteins were titrated against EloBC. The raw data are shown on top, the heat integration at the bottom. The resulting Kd is given for each construct, except for the ΔSLQ protein, for which no binding was observed. (D) Thermodynamic analysis of the ITC binding assay. The binding free energy (ΔG), enthalpy (ΔH) and entropy (ΔS) are plotted for the Vif fusions proteins binding to EloBC. The ΔSLQ protein is not shown as no binding was observed.