Figure 4. The catalytic motif CX 5 R of... | Rockefeller University Press

Figure 4.

The catalytic motif CX5R of TMEM55B is responsible for interacting with Tex2. (A) Domain organization of TMEM55B. (B) Representative images of a COS7 cell expressing Halo-TMEM55B-∆TM (magenta), ER-GFP (green), and Lamp1-mCh (blue) with insets. (C–I) Representative images of COS7 cells expressing either Halo-TMEM55B-∆TM (magenta; C), Halo-TMEM55B-∆NT (75-284aa; D), Halo-TMEM55B-TM (E), Halo-TMEM55B-∆PTase (F), Halo-TMEM55B-∆X5R (G), Halo-PP4P1-C140W (H), and Halo-PP4P1-C140S (I). Yellow arrows indicated TMEM55B-positive LE/lys with GFP-Tex2 enrichments; while red arrows denoted TMEM55B-positive LE/lys without GFP-Tex2 enrichments. (J) Pearson’s correlation coefficient of TMEM55B proteins vs. Tex2; Halo-TMEM55B-∆TM in absence of Tex2 (9 cells); and Halo-TMEM55B-∆TM (9 cells); Halo-TMEM55B-∆1-74 (10 cells); Halo-TMEM55B-∆1-163 (9 cells); Halo-TMEM55B-∆PTase (9 cells); Halo-TMEM55B-∆CX5R (9 cells); Halo-PP4P1-C140W (10 cells); and Halo-PP4P1-C140S (10 cells) upon Tex2 overexpression in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (K) Representative images of either control (left panel) or Tex2 KO-1 (right panel) HeLa cells expressing GFP-TMEM55B (green) and Lamp1-mCh (magenta) with insets. (L) Pearson’s correlation coefficient of TMEM55B vs. Lamp1 in either control (21 cells) or Tex2 KO-1 (20 cells) HeLa cells in three independent experiments. Two-tailed unpaired Student’s t test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in B–I and K.

The catalytic motif CX ₅ R of TMEM55B is responsible for interacting with Tex2. (A) Domain organization of TMEM55B. (B) Representative images of a COS7 cell expressing Halo-TMEM55B-∆TM (magenta), ER-GFP (green), and Lamp1-mCh (blue) with insets. (C–I) Representative images of COS7 cells expressing either Halo-TMEM55B-∆TM (magenta; C), Halo-TMEM55B-∆NT (75-284aa; D), Halo-TMEM55B-TM (E), Halo-TMEM55B-∆PTase (F), Halo-TMEM55B-∆X₅R (G), Halo-PP4P1-C140W (H), and Halo-PP4P1-C140S (I). Yellow arrows indicated TMEM55B-positive LE/lys with GFP-Tex2 enrichments; while red arrows denoted TMEM55B-positive LE/lys without GFP-Tex2 enrichments. (J) Pearson’s correlation coefficient of TMEM55B proteins vs. Tex2; Halo-TMEM55B-∆TM in absence of Tex2 (9 cells); and Halo-TMEM55B-∆TM (9 cells); Halo-TMEM55B-∆1-74 (10 cells); Halo-TMEM55B-∆1-163 (9 cells); Halo-TMEM55B-∆PTase (9 cells); Halo-TMEM55B-∆CX₅R (9 cells); Halo-PP4P1-C140W (10 cells); and Halo-PP4P1-C140S (10 cells) upon Tex2 overexpression in three independent experiments. Ordinary one-way ANOVA with Tukey’s multiple comparisons test. Mean ± SD. (K) Representative images of either control (left panel) or Tex2 KO-1 (right panel) HeLa cells expressing GFP-TMEM55B (green) and Lamp1-mCh (magenta) with insets. (L) Pearson’s correlation coefficient of TMEM55B vs. Lamp1 in either control (21 cells) or Tex2 KO-1 (20 cells) HeLa cells in three independent experiments. Two-tailed unpaired Student’s t test. Mean ± SD. Scale bar, 10 μm in the whole cell images and 2 μm in the insets in B–I and K.