FIGURE 1.

Orientation of reconstituted Glt_Ph. A, fluorescein-5-maleimide labeling and Coomassie Blue staining of Glt_Ph mutants, Cys-less (C321S), and with a single cysteine on the extracellular (G34C) and intracellular (S72C) faces of the protein, in liposomes before (L) and after Triton X-100 solubilization (S) of liposomes. Protein had been reconstituted into vesicles swollen with Triton X-100 at a ratio 0.5 (w/w) detergent to lipid. Top panel represents gel imaged on a UV light box; bottom panel represents the same gel, subsequently stained with Coomassie Blue and imaged with visible transillumination. B, swelling of vesicles measured by light scattering of vesicles (absorbance at 540 nm) as a function of Triton X-100 to lipid ratio. C, gels of labeling experiments as described in A but reconstituted at Triton/lipid ratios of 0.25 (w/w) or in the absence of Triton X-100 (0 w/w).