Immunological Quantitation and Localization of ACAT-1 and ACAT-2 in Human Liver and Small Intestine

作者：Catherine C.Y. Chang, Naomi Sakashita, Kim Ørnvold, Oneil Lee, Ellen T. Chang, Ruhong Dong, Song Lin, Chi‐Yu Gregory Lee, Stephen C. Strom, Randeep Kashyap, John J. Fung, Robert V. Farese, Jean‐François Patoiseau, André Delhon, Ta‐Yuan Chang · 发表于：Journal of Biological Chemistry · 年份：2000 · DOI：10.1074/jbc.m003927200 · 被引用次数：234 · 研究领域：Cholesterol and Lipid Metabolism、Drug Transport and Resistance Mechanisms、Peroxisome Proliferator-Activated Receptors

By using specific anti-ACAT-1 antibodies in immunodepletion studies, we previously found that ACAT-1, a 50-kDa protein, plays a major catalytic role in the adult human liver, adrenal glands, macrophages, and kidneys but not in the intestine. Acyl-coenzyme A:cholesterol acyltransferase (ACAT) activity in the intestine may be largely derived from a different ACAT protein. To test this hypothesis, we produced specific polyclonal anti-ACAT-2 antibodies that quantitatively immunodepleted human ACAT-2, a 46-kDa protein expressed in Chinese hamster ovary cells. In hepatocyte-like HepG2 cells, ACAT-1 comprises 85-90% of the total ACAT activity, with the remainder attributed to ACAT-2. In adult intestines, most of the ACAT activity can be immunodepleted by anti-ACAT-2. ACAT-1 and ACAT-2 do not form hetero-oligomeric complexes. In differentiating intestinal enterocyte-like Caco-2 cells, ACAT-2 protein content increases by 5-10-fold in 6 days, whereas ACAT-1 protein content remains relatively constant. In the small intestine, ACAT-2 is concentrated at the apices of the villi, whereas ACAT-1 is uniformly distributed along the villus-crypt axis. In the human liver, ACAT-1 is present in both fetal and adult hepatocytes. In contrast, ACAT-2 is evident in fetal but not adult hepatocytes. Our results collectively suggest that in humans, ACAT-2 performs significant catalytic roles in the fetal liver and in intestinal enterocytes.