Crystal structure of the human liver X receptor beta ligand-binding domain in complex with a synthetic agonist.

LXRbeta belongs to the nuclear hormone receptor superfamily of ligand-activated transcription factors. Its natural ligands are supposed to be oxidised derivatives of cholesterol. Stimulation of LXRbeta by agonists activates a number of genes that are involved in the regulation of lipid metabolism and cholesterol efflux from cells. Therefore, LXRbeta may represent a novel therapeutic target for the treatment of dyslipidemia and atherosclerosis.Here, we report the X-ray crystal structure of the LXRbeta ligand-binding domain in complex with a synthetic agonist, T-0901317. This compound occupies the ligand-binding pocket of the receptor, forms numerous lipophilic contacts with the protein and one crucial hydrogen bond to His435 and stabilises the agonist conformation of the receptor ligand-binding domain. The recruitment of the AF2-region of the protein is not achieved via direct polar interactions of the ligand with protein side-chains of this helical segment, but rather via few hydrophobic contacts and probably more importantly via indirect effects involving the pre-orientation of side-chains that surround the ligand-binding pocket and form the interface to the AF2-helix. On the basis of these results we propose a binding mode and a mechanism of action for the putative natural ligands, oxidised derivatives of cholesterol.

Scavenger receptor BI (SR-BI) mediates a higher selective cholesteryl ester uptake from LpA-I compared with LpA-I:A-II lipoprotein particles.

Scavenger receptor class B, type I (SR-BI) mediates the selective uptake of high-density lipoprotein- (HDL-) associated cholesteryl esters (CE), i.e. lipid uptake independent from HDL holo-particle internalisation. This pathway contributes to the HDL-mediated CE delivery to the liver. From human plasma HDL, two major lipoprotein subfractions can be isolated: one contains apolipoprotein (apo) A-I and apo A-II (LpA-I:A-II) as dominant protein components, whereas in the other population apo A-II is absent (LpA-I). In this investigation the role of SR-BI in selective CE uptake from HDL, LpA-I and LpA-I:A-II was explored. HDL(3) (d=1.125-1.21 g/ml), LpA-I and LpA-I:A-II were isolated from human plasma and radiolabeled in the protein (125I) as well as in the CE moiety ([3H]). Baby hamster kidney (BHK) cells were stably transfected with the full-length human SR-BI cDNA and these cells demonstrate SR-BI expression in immunoblots. In contrast, no SR-BI protein was detectable in control BHK cells (vector). To investigate lipoprotein uptake, cells incubated (37 degrees C, 4 h) in medium containing radiolabeled HDL(3), LpA-I or LpA-I:A-II and finally cellular tracer content was determined. For both types of BHK cells, the rate of apparent lipoprotein particle uptake according to the lipid tracer ([3H]) was in substantial excess over that due to the protein tracer (125I) demonstrating selective CE uptake ([3H]-(125)I) from HDL(3), LpA-I and LpA-I:A-II. SR-BI expression increased cellular selective CE uptake from labeled HDL(3) up to 24-fold. In BHK cells without SR-BI expression, selective CE uptake was higher from LpA-I compared with LpA-I:A-II. Analogously, in BHK cells with SR-BI expression, the rate of selective CE uptake was higher from LpA-I compared with LpA-I:A-II. In summary, SR-BI significantly increases selective CE uptake from HDL(3), LpA-I and LpA-I:A-II. Concerning HDL subfractions, the rate of SR-BI-mediated selective CE uptake is greater from LpA-I compared with LpA-I:A-II and this result suggests that SR-BI preferentially facilitates the CE transfer from LpA-I lipoprotein particles to cells.