Insulin degradation. XXIII. Distribution of glutathione-insulin transhydrogenase in isolated rat hepatocytes as studied by immuno-ferritin and electron microscopy.
The distribution of glutathione-insulin transhydrogenase (glutathione: protein-disulphide oxidoreductase, EC 1.8.4.2) in isolated rat hepatocytes that had been first treated with rabbit antiserum against purified rat liver transhydrogenase and then with ferritin-conjugated goat anti-rabbit gamma-globulin was examined by electron microscopy. In cells with intact plasma membrane, the immunoferritin labeling of glutathione-insulin transhydrogenase was observed on a few external microvillous projections at the outside of the cell. In cells with breaks in the plasma membrane, the immunoferritin labeling appeared extensively on smooth vesicles just inside the plasma membrane and on smooth endoplasmic reticulum extending to and including the outer nuclear membrane, in addition to the external microvillous projections. There was some immunoferritin labeling on rough endoplasmic reticulum and on the inner surface of the plasma membrane. The mitochondria and the outer surface of the plasma membrane of the cell did not show the ferritin labeling. Control parallel samples in which the antiserum was substituted with normal (i.e. non-immune) serum or with neutralized antiserum (prepared by absorption with the transhydrogenase) showed little or no immunoferritin labeling. These results are consistent with the idea that gluthalione-insulin transhydrogenase probably synthesized in the endoplasmic reticulum and that the transhydrogenase accessible to cell surface (or found in the isolated plasma membrane preparations) probably represents a functional continuity between the endoplasmic reticulum and the plasma membrane.
D000818:Animals; D002462:Cell Membrane / Q000201:enzymology; D004721:Endoplasmic Reticulum / Q000201:enzymology; D007158:Immunologic Techniques; D007328:Insulin / Q000378:metabolism*; D008099:Liver / Q000201:enzymology* / Q000648:ultrastructure; D008871:Microvilli / Q000201:enzymology; D010088:Oxidoreductases / Q000378:metabolism*; D011490:Protein Disulfide Reductase (Glutathione) / Q000378:metabolism*; D051381:Rats
D007328:Insulin; D010088:Oxidoreductases; D011490:Protein Disulfide Reductase (Glutathione)