Regulation of acyl-coenzyme A:cholesterol acyltransferase (ACAT) synthesis, degradation, and translocation by high-density lipoprotein(2) at a low concentration.
(,Although plasma HDL(2) cholesterol concentration stands in inverse relation to risk for atherosclerotic disease, little is known about the mechanism of the apparent cardioprotection. In mouse P388D1 macrophages, HDL(2) at a low concentration (< or = 40 microg/mL) inhibits macrophage acyl-coenzyme A:cholesterol acyltransferase (ACAT), the enzyme that catalyzes esterification of intracellular cholesterol. The effects of HDL(2) on ACAT synthesis, degradation, and intracellular translocation were investigated in mouse P388D1 macrophages. HDL(2) at a low concentration enhanced ACAT synthesis but not total ACAT mass. Immunocytochemical studies showed that in the absence of lipoproteins, ACAT associated primarily with the perinuclear region of the cell. The addition of HDL(2), however, induced the transfer of ACAT to vesicular structures and the cell periphery adjacent to the plasma membrane. Subfractionation combined with immunoprecipitation complemented these observations and showed that HDL(2) promoted the transfer of ACAT to the plasma membrane fraction. Brefeldin A, which inhibits vesicular protein transport from the endoplasmic reticulum to the Golgi compartment in mammalian cells, blocked ACAT translocation and partially restored ACAT activity. These results suggest that HDL(2) is an initiating factor in a signal transduction pathway that leads to intracellular ACAT translocation and inactivation.
Li
L
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Department of Medicine, Baylor College of Medicine, and The Methodist Hospital, Houston, TX 77030, USA.
Pownall
H J
HJ
D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D002250:Carbon Radioisotopes; D002458:Cell Fractionation; D002460:Cell Line; D002462:Cell Membrane / Q000201:enzymology; D002788:Cholesterol Esters / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004721:Endoplasmic Reticulum / Q000201:enzymology; D004789:Enzyme Activation / Q000187:drug effects; D005455:Fluorescent Antibody Technique; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D006056:Golgi Apparatus / Q000201:enzymology; D007150:Immunohistochemistry; D008075:Lipoproteins, HDL / Q000494:pharmacology*; D008264:Macrophages / Q000187:drug effects* / Q000201:enzymology; D051379:Mice; D009685:Nuclear Envelope / Q000201:enzymology; D015398:Signal Transduction / Q000187:drug effects; D002785:Sterol O-Acyltransferase / Q000032:analysis / Q000096:biosynthesis / Q000378:metabolism*
D002250:Carbon Radioisotopes; D002788:Cholesterol Esters; D008075:Lipoproteins, HDL; D020126:Brefeldin A; D002785:Sterol O-Acyltransferase