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Figure 4


Fig. 4. Binding of the acidic peptide region of Furin by Mint3. (A) Schematic diagrams of Furin and its various deletion mutants. The binding abilities of GST-tagged fusion proteins to Mint3 are summarized to the right. ND, not determined. (B) The association of the cytoplasmic domain of Furin with Mint3. GST fusion proteins of the catalytic domain (GST-CA), the P-domain (GST-P) and the cytoplasmic domain (GST-CP) of Furin were immobilized on glutathione-Sepharose beads and incubated with lysates from HeLa cells. Bound proteins were eluted, separated by SDS-PAGE and visualized as indicated. TCL (total cell lysate) is shown as 5% of input. (C) Schematic representation of GST fusion proteins with various mutations of the Furin cytoplasmic domain. Deleted regions are marked by a dotted line and amino acid substitutions are labeled in red. Binding motifs are bold and underlined. The binding abilities of GST-tagged fusion proteins to Mint3 are summarized to the right. (D) The mutants with acidic peptide deletion ({Delta}769-780) fail to bind Mint3 efficiently, and the mutants with LI/AN Y/A substitutions decrease the binding affinity between Furin and Mint3.





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