FIGURE 1.

Electrophoresis of GGT from human kidney and liver microsomes. A, native gel analysis of GGT. Triton X-100-solubilized microsomes from normal human kidney (lanes 1–3 and 7–9) or liver (lanes 5 and 6 and 11 and 12) tissue were incubated at 37 °C with papain alone (lanes 1–6) or with papain and clostridial neuraminidase (lanes 7–12) and resolved in parallel on 10% nondenaturing polyacrylamide gels. GGT was localized in situ with the histochemical stain for GGT activity (red precipitate). B, SDS-PAGE analysis of intact and deglycosylated GGT. Non-papain-treated samples from human kidney (lanes 1 and 2 and 6 and 7) and liver (lanes 3 and 4 and 8 and 9) were denatured and incubated in the absence (lanes 1–4) or presence (lanes 6–9) of PNGase F and resolved on an 8% (upper panel) or 10% (lower panel) SDS-polyacrylamide gel. Resolved proteins were electroblotted onto nitrocellulose and Western blotted using antibodies against the large and small subunits of GGT. Positions of molecular mass (M) markers are indicated.