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. 2009 Jul 13;186(1):41–55. doi: 10.1083/jcb.200902110

Figure 7.

Figure 7.

Mutation of the predicted PDZ1 ligand-binding groove blocks clustering. GRASP sequences are shown aligned at the position predicted by the TASSERlite modeling program (Zhang and Skolnick, 2004) to correspond to the second α-helix of the first PDZ domain (A). As illustrated in the diagram (B), this helix is oriented in the model such that two leucines (L58, L59) face the binding pocket formed between the α-helix and a β-strand. HeLa (C) or mfn−/− (D) cells expressing G65LL58,59SS-GFP-ActA were analyzed using GFP fluorescence after BFA treatment (bar = 10 µm) and the radial profile plot for HeLa cells (E) is shown (n = 3, ±SEM, >15 cells/experiment). HeLa cells expressing GalNAcT2-GFP and transfected with GRASP65 siRNA and either no vector (F–H), G65-myc (I–K), or G65LL58,59SS-myc (L–N) were analyzed to assess Golgi morphology (green) and replacement construct expression (red). Bar, 10 µm. Percentage of cells transfected with G65-myc or G65LL58,59SS-myc exhibiting a fragmented Golgi after knockdown with GRASP65 siRNAs (±SEM, n = 3, >50 cells each; *, P < 0.0001).