Immunolocalization of acyl-coenzyme A:cholesterol O-acyltransferase in macrophages
| Autor: | Nadia Khelef, Ta-Yuan Chang, Nanda Beatini, Robert V. Farese, Xavier Buton, Ira Tabas, Hongxing Wang, Frederick R. Maxfield, Vardiella L. Meiner |
|---|---|
| Rok vydání: | 1998 |
| Předmět: |
Sterol O-acyltransferase
Fluorescent Antibody Technique Ribophorin Lipoproteins VLDL Biochemistry Cell membrane Mice medicine Animals Nuclear membrane Molecular Biology Foam cell Mice Inbred ICR biology Esterification Endoplasmic reticulum Macrophages Cell Membrane Cell Biology Subcellular localization Cell biology Mice Inbred C57BL medicine.anatomical_structure Cholesterol Cytoplasm biology.protein Female Sterol O-Acyltransferase |
| Zdroj: | The Journal of biological chemistry. 273(18) |
| ISSN: | 0021-9258 |
| Popis: | Macrophages in atherosclerotic lesions accumulate large amounts of cholesteryl-fatty acyl esters ("foam cell" formation) through the intracellular esterification of cholesterol by acyl-coenzyme A:cholesterol O-acyltransferase (ACAT). In this study, we sought to determine the subcellular localization of ACAT in macrophages. Using mouse peritoneal macrophages and immunofluorescence microscopy, we found that a major portion of ACAT was in a dense reticular cytoplasmic network and in the nuclear membrane that colocalized with the luminal endoplasmic reticulum marker protein-disulfide isomerase (PDI) and that was in a similar distribution as the membrane-bound endoplasmic reticulum marker ribophorin. Remarkably, another portion of the macrophage ACAT pattern did not overlap with PDI or ribophorin, but was found in as yet unidentified cytoplasmic structures that were juxtaposed to the nucleus. Compartments containing labeled beta-very low density lipoprotein, an atherogenic lipoprotein, did not overlap with the ACAT label, but rather were embedded in the dense reticular network of ACAT. Furthermore, cell-surface biotinylation experiments revealed that freshly harvested, non-attached macrophages, but not those attached to tissue culture dishes, contained approximately 10-15% of ACAT on the cell surface. In summary, ACAT was found in several sites in macrophages: a cytoplasmic reticular/nuclear membrane site that overlaps with PDI and ribophorin and has the characteristics of the endoplasmic reticulum, a perinuclear cytoplasmic site that does not overlap with PDI or ribophorin and may be another cytoplasmic structure or possibly a unique subcompartment of the endoplasmic reticulum, and a cell-surface site in non-attached macrophages. Understanding possible physiological differences of ACAT in these locations may reveal an important component of ACAT regulation and macrophage foam cell formation. |
| Databáze: | OpenAIRE |
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