Targeting and Function of the Mitochondrial Fission Factor GDAP1 Are Dependent on Its Tail-Anchor (original) (raw)
Figure 1
GDAP1 membrane topology.
(A) Graphic of experimental strategy. (B) Western blot of mitochondria-enriched fractions derived from GDAP1-FLAG-transfected HeLa cells treated with proteinase K for 30 minutes at 4°C with increasing digitonin concentrations. Without digitonin, the cytosolic N-terminal part of GDAP1-FLAG is degraded (arrow) and only the C-terminal 5 kDa fragment is detected (arrowhead). After membrane permeabilization, the C-terminal fragment is gradually degraded confirming its location in the IMS. The IMS protein Opa1 served as control. (C) Confocal immunofluorescence microscopy of transfected COS-7 cells expressing GDAP1-FLAG (a–d). Co-stainings: MitoTracker (mitochondria). Bars, 10 µm.