Fig 3.

Requirement of JAMM domain of JAB1 for its binding with Δ2 LHBS in the presence of Zn²⁺ ions. (A) Schematic representations showing the partial deletion constructs of JAB1 used in the present study. The wild-type (FL) and mutant JAB1 proteins (Δ1-52, Δ251-334, and ΔJAMM) were N-terminal fused to GST. (B) GST pulldown assays to detect the binding of Δ2 LHBS to JAB1 in the presence of (2 μM, + Zn²⁺) or absence of Zn²⁺ ions, followed with Western blotting (WB) with antibodies that recognize GST (top panel) and LHBS (middle panel). The bottom panel shows the input amounts of the JAB1 proteins to the GST pull-down reactions, detected by using Coomassie blue staining. (C) Protein binding of Δ2 LHBS to JAB1 and its H138Y mutant in the presence of Zn²⁺ ions, detected by using GST pulldown assays, followed by WB to detect LHBS (top panel) and GST (bottom panel).