BIOSYNTHESIS AND SUBCELLULAR LOCATION OF H-2 HISTOCOMPATIBILITY ANTIGENS OF MOUSE LIVER
Abstract
The intracellular transport and subcellular distribution of H-2 histocompatibility antigens of mouse liver were studied. Specific immunoprecipitation and immunocytochemistry were used to localize H-2 histocompatibility antigens on membranes of the endoplasmic reticulum, Golgi apparatus and cell surface. A fractionation procedure was developed and characterized for preparation of purified membranes of endoplasmic reticulum, Golgi apparatus and plasma membranes from single homogenates of mouse liver. These cell components then were used to determine the subcellular localization of H-2 histocompatibility antigens and delineate the intracellular transport route of these antigens from within the cell to the cell surface. H-2 histocompatibility antigens were found to be concentrated on plasma membranes and also associated with membranes of the endoplasmic reticulum and Golgi apparatus. H-2 histocompatibility antigens migrated from their site of synthesis on the endoplasmic reticulum to the Golgi apparatus within 15 min. Migration of H-2 antigens from the Golgi apparatus to the cell surface took place within 40 min. H-2 antigens once associated with the plasma membrane turned over very slowly. Thus, using specific immunoprecipitation of H-2 histocompatibility antigens and purified cell fractions it was possible to demonstrate the intracellular transport (flow) of a single purified membrane glycoprotein (H-2 antigen) from its site of synthesis within the cell to the cell surface. The intracellular migration of H-2 histocompatibility antigens began after insertion into membranes of the endoplasmic reticulum and was followed by passage through the Golgi apparatus and final association with the plasma membrane.
Degree
Ph.D.
Subject Area
Biology
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