Figure 2: Interaction between VPS29 and TBC1d5.
From: Structural and mechanistic insights into regulation of the retromer coat by TBC1d5
(a) Ribbon diagram of the VPS29/TBC1d5-Ins1 complex (VPS29: green; Ins1; yellow). (b) Structural comparison of VPS29/TBC1d5-Ins1 and VPS29/VPS35C. VPS29 is shown as same orientation in two structures and is 90° rotation about a horizontal axis from a. (VPS35C: magenta). (c) GRASP surface charge distribution (blue for positive, red for negative, white for neutral). The Ins1 is shown as stick models. The structure is shown as same orientation as a. (d,e) VPS29-TBC1d5-Ins1 interactions in detail. Critical VPS29 and TBC1d5-Ins1 residues are shown in stick modes. (f,g) Coomassie blue stained SDS–PAGE gels of eluted proteins are shown. In f, immobilized GST–TBC1d5TBC WT but not mutants (ΔIns1, NPL/EEE, N140A, P141A, L142A, L142E, L142W and W150A) selectively retained CSC. In g, mutation on VPS29 surface residues had various effects on the interaction between GST–TBC1d5TBC and VPS35C/VPS29. (h) TBC1d5TBC WT, but not ΔIns1, NPL/EEE, L142E and W150A, co-immunoprecipitated with VPS35. Hela cells were transfected with yellow fluorescent protein (YFP) or various YFP–TBC1d5 (green) and immunoprecipitate with anti-VPS35 or anti-HA antibodies. Lysates were immunoblotted as a control.
