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. Author manuscript; available in PMC: 2018 Dec 10.
Published in final edited form as: EMBO Rep. 2018 Nov 7;19(12):e45918. doi: 10.15252/embr.201845918

Figure 5. Rab22A regulates membrane association of BLOC-1 and BLOC-2, and forms a complex with BLOC-1-BLOC-2-KIF13A.

Figure 5

A Subcellular membrane fractionation of control and Rab22A-, BLOC-1-, BLOC-2-knockdown HeLa cells and probed the fractions for pallidin, dysbindin and muted. Red coloured box emphasizes BLOC-1 membrane association in the respective cell types.

B, C Membrane-cytosol fractionation of homogenates from HeLa cells (B) or melanocytes (C) as indicated. Protein band intensities were quantified and indicated the percentage membrane association on the gels.

D, E Pull-down of His-Rab22AWT using HeLa (D) or melanocyte (E) lysate. In (E), the beads were preloaded with GTPγS or GDP.

F Pull-down of different His-KIF13A domains using HeLa cell lysate.

G Model illustrating the recruitment (left) and association (middle) of Rab22A, BLOC-1 and BLOC-2 in a sequential manner onto the membrane buds of E/SEs followed by interaction with KIF13A motor. Rab22A-BLOC-1-BLOC-2 complex possibly extends the membrane buds into RE tubules with KIF13A motor (right) movement on microtubules.

Data information: In (A–F), * indicates non-specific bands. In (D–F), the bead-bound His-Rab22A/His-KIF13A domains were shown on the Coomassie-stained gels separately or in Fig 1F.