Fig. 7: GRASP55-PDZ1 interacts with ATGL-patatin.
From: Grasp55−/− mice display impaired fat absorption and resistance to high-fat diet-induced obesity
a–d Coimmunoprecipitation experiments with anti-Myc antibodies were performed in Caco-2 cells. For the induction of ATGL protein expression, some cells were treated with 400 μM oleic acid for 16 h. A representative coimmunoprecipitation assay using GRASP55 with a COOH-terminal Myc-tag (GRASP55-Myc) is shown in (a), and the results of multiple experiments are summarized in (b, n = 3). A representative coimmunoprecipitation assay using ATGL with a COOH-terminal Myc-tag (ATGL-Myc) is shown in (c), and the results of multiple experiments are summarized in (d) (n = 3). Aldolase A was used as a cytosolic protein loading control. e, f Pull-down assays were performed with GRASP55 and ATGL fragments. The domain structures of His6-tagged GRASP55 and GST-tagged ATGL constructs used in this study are shown in (e), and a representative result from the pull-down assay is shown in (f). Expression of control GST and each GST-fusion protein is visualized by Ponceau S staining (f lowermost panel). An asterisk indicates the band of each GST-tagged protein. One microgram of each His6-tagged protein was loaded as an input control. The PDZ1 domain of GRASP55 interacted strongly with the ATGL-patatin domain. Three independent experiments showed similar results. Unprocessed blots can be found in Supplementary Fig. 22. Data are shown as mean ± SEM. n.s.: not significant, *p < 0.05, **p < 0.01. All p values were calculated by paired two-tailed Student’s t tests. Source data are provided as a Source Data file.
