Fig. 1. Effect of metabolic stress on surface labelling of GLUT1 in Clone 9 cells
by a membrane-impermeant photoaffinity reagent. Clone 9 cells were incubated
with or without 5 mM sodium azide for 30 minutes, as indicated, then
photolabelled with 500 µM Bio-LC-ATB-BMPA in the absence or presence of 400
mM glucose (lane 3) or 20 µM cytochalasin B (lane 4). A control sample
(lane 5) was also UV-irradiated in the absence of Bio-LC-ATB-BMPA. Following
cell lysis with 1% TX-100, samples of total lysate proteins (a) and of
biotinylated cell-surface proteins isolated by adsorption to streptavidin
agarose (b) were subjected to western blotting to detect GLUT1, as described
in the Materials and Methods. The mobilities of the molecular mass markers are
indicated on the left.
