Cholesterol Journal Articles

Record 11961 to 11980

Structure of a cholesterol-binding protein deficient in Niemann-Pick type C2 disease
Friedland, N., H. L. Liou, et al. (2003), Proc Natl Acad Sci U S A 100(5): 2512-7.
Abstract: Niemann-Pick disease type C2 (NP-C2) is a fatal hereditary disease characterized by accumulation of low-density lipoprotein-derived cholesterol in lysosomes. Here we report the 1.7-A resolution crystal structure of the cholesterol-binding protein deficient in this disease, NPC2, and the characterization of its ligand binding properties. Human NPC2 binds the cholesterol analog dehydroergosterol with submicromolar affinity at both acidic and neutral pH. NPC2 has an Ig-like fold stabilized by three disulfide bonds. The structure of the bovine protein reveals a loosely packed region penetrating from the surface into the hydrophobic core that forms adjacent small cavities with a total volume of approximately 160 A(3). We propose that this region represents the incipient cholesterol-binding site that dilates to accommodate an approximately 740-A(3) cholesterol molecule.

Structure of a cholesterol-binding, thiol-activated cytolysin and a model of its membrane form
Rossjohn, J., S. C. Feil, et al. (1997), Cell 89(5): 685-92.
Abstract: The mechanisms by which proteins gain entry into membranes is a fundamental problem in biology. Here, we present the first crystal structure of a thiol-activated cytolysin, perfringolysin O, a member of a large family of toxins that kill eukaryotic cells by punching holes in their membranes. The molecule adopts an unusually elongated shape rich in beta sheet. We have used electron microscopy data to construct a detailed model of the membrane channel form of the toxin. The structures reveal a novel mechanism for membrane insertion. Surprisingly, the toxin receptor, cholesterol, appears to play multiple roles: targeting, promotion of oligomerization, triggering a membrane insertion competent form, and stabilizing the membrane pore.

Structure of cholesterol/ceramide monolayer mixtures: implications to the molecular organization of lipid rafts
Scheffer, L., I. Solomonov, et al. (2005), Biophys J 88(5): 3381-91.
Abstract: The structure of monolayers of cholesterol/ceramide mixtures was investigated using grazing incidence x-ray diffraction, immunofluorescence, and atomic force microscopy techniques. Grazing incidence x-ray diffraction measurements showed the existence of a crystalline mixed phase of the two components within a range of compositions of cholesterol/ceramide between 100:0 and 67:33. The mixed phase coexists with the ceramide crystalline phase in the range of compositions between 50:50 and 30:70; between 30:70 and 0:100 only the highly crystalline phase of ceramide was detected. The latter was determined and modeled. Immunolabeling was performed with an antibody specific to the cholesterol monohydrate crystalline arrangement. The antibody recognizes crystalline cholesterol monolayers, but does not interact with crystalline ceramide. Immunofluorescence and atomic force microscopy data show that in uncompressed ceramide monolayers, the highly crystalline phase coexists with a disordered loosely packed phase. In contrast, no disordered phase coexists with the new crystalline mixed phase. We conclude that the new mixed phase represents a stable homogeneous arrangement of cholesterol with ceramide. As ceramide incorporates the lipid backbone common to all sphingolipids, this arrangement may be relevant to the understanding of the molecular organization of lipid rafts.

Structure of cholesterol-containing particles accumulating in atherosclerotic lesions and the mechanisms of their derivation
Hoff, H. F. and G. Hoppe (1995), Curr Opin Lipidol 6(5): 317-25.
Abstract: Structural and chemical modifications of plasma lipoproteins retained in atherosclerotic lesions, especially LDL, are a characteristic of atherogenesis. The major cholesterol-containing structures believed to be derived primarily from LDL are monomeric or aggregated native or modified LDL particles, cholesteryl ester droplets, liposomes rich in unesterified cholesterol, and ceroid-lipofuscin. They are suggested to be formed primarily from LDL by combinations of oxidation, hydrolysis by proteases and esterases, fusion of neutral lipid components, and covalent interactions between lipid and protein components of oxidized LDL in lysosomes. Although many of these structures appear to be refractory to removal by reverse cholesterol transport mechanisms, they may possess functional properties that still need to be elucidated.

Structure of dipalmitoylphosphatidylcholine/cholesterol bilayer at low and high cholesterol concentrations: molecular dynamics simulation
Smondyrev, A. M. and M. L. Berkowitz (1999), Biophys J 77(4): 2075-89.
Abstract: By using molecular dynamics simulation technique we studied the changes occurring in membranes constructed of dipalmitoylphosphatidylcholine (DPPC) and cholesterol at 8:1 and 1:1 ratios. We tested two different initial arrangements of cholesterol molecules for a 1:1 ratio. The main difference between two initial structures is the average number of nearest-neighbor DPPC molecules around the cholesterol molecule. Our simulations were performed at constant temperature (T = 50 degrees C) and pressure (P = 0 atm). Durations of the runs were 2 ns. The structure of the DPPC/cholesterol membrane was characterized by calculating the order parameter profiles for the hydrocarbon chains, atom distributions, average number of gauche defects, and membrane dipole potentials. We found that adding cholesterol to membranes results in a condensing effect: the average area of membrane becomes smaller, hydrocarbon chains of DPPC have higher order, and the probability of gauche defects in DPPC tails is lower. Our results are in agreement with the data available from experiments.

Structure of stratum corneum lipids characterized by FT-Raman spectroscopy and DSC. III. Mixtures of ceramides and cholesterol
Wegener, M., R. Neubert, et al. (1997), Chem Phys Lipids 88(1): 73-82.
Abstract: The thermotropic and lyotropic phase behaviour of mixtures of ceramides type IV and cholesterol was investigated using Fourier transform (FT) Raman spectroscopy and differential scanning calorimetry (DSC). In the dry and in the fully hydrated state of these mixtures the DSC-curves exhibit an eutectic melting followed by the melting of the residual solid component. The Raman spectrum of the mixtures is complex, nevertheless, the appearance of the conformationally dependent bands indicates the ordered structure of the hydrocarbon chains. The temperature dependence of the conformationally sensitive bands in the CH2 stretching region (2800-2975 cm-1) and in the chain C-C stretching region (1050-1150 cm-1) was used to estimate the degree of order in terms of the relative population of trans and gauche conformers. The spectrum of the pure cholesterol shows only a weak temperature dependence in the CH2 stretching region and, therefore, the decrease of the intensity of the asymmetric CH2 stretching mode at 2880 cm-1 can be attributed to the melting of the alkyl chains of ceramides. The temperature and width of the phase transition, derived from Raman data, are similar to those of the DSC study.

Structure of the human steroidogenic acute regulatory protein (StAR) gene: StAR stimulates mitochondrial cholesterol 27-hydroxylase activity
Sugawara, T., D. Lin, et al. (1995), Biochemistry 34(39): 12506-12.
Abstract: Steroidogenic acute regulatory protein (StAR) plays a key role in steroid hormone synthesis by enhancing the metabolism of cholesterol into pregnenolone. We determined the organization of the StAR structural gene, mapped to 8p11.2. The gene spans 8 kb and consists of seven exons interrupted by six introns. The 1.3 kb of DNA upstream from the transcription start site directed expression of a luciferase reporter gene in mouse Y-1 adrenal cortical tumor cells but not in BeWo choriocarcinoma cells. Reporter gene expression in the Y-1 cells was increased more than 2-fold by 8-Br-cAMP, indicating that the 1.3 kb DNA fragment contains sequences that confer tissue-specific expression and cAMP regulation. The sequence of a related StAR pseudogene, mapped to chromosome 13, lacks introns and has an insertion, numerous substitutions, and deletions. Expression of StAR in COS-1 cells cotransfected with cholesterol 27-hydroxylase (P450c27) and adrenodoxin resulted in a 6-fold increase in formation of 3 beta-hydroxy-5-cholestenoic acid, demonstrating that StAR's actions are not specific to steroidogenesis but extend to other mitochondrial cholesterol-metabolizing enzymes.

Structure-activity relationships of cytotoxic cholesterol-modified DNA duplexes
Reed, M. W., E. A. Lukhtanov, et al. (1995), J Med Chem 38(22): 4587-96.
Abstract: Short DNA duplexes with cholesterol linked at the 3'-terminus of each strand have unique, selective cytotoxic properties. The structural requirements for biological activity were explored through chemical synthesis of analogs and testing in cultured hepatoma cells. Effects of modifications to the sequence, backbone, 3'-sterol, 3'-linker, and 5'-terminus were evaluated. Self-complementary 3'-modified oligodeoxynucleotide (ODN) 10-mers were prepared from solid supports bearing the modification and linker of interest. Any changes to the normal phosphodiester backbone were poorly tolerated. The presence of cholesterol or a closely related sterol was an absolute requirement for activity. The length and position of attachment of the linker to cholesterol was important, with longer linkers showing reduced activity. Large, lipophilic groups at the 5'-terminus gave reduced cytotoxicity and poor solubility properties. The short length and unique structure of these ODNs allowed efficient automated synthesis on a 400 mumol scale and simplified purification.

Structure-activity relationships of N-(3,5-dimethoxy-4-n-octyloxycinnamoyl)-N'-(3,4-dimethylphenyl)piperazine and analogues as inhibitors of acyl-CoA: cholesterol O-acyltransferase
Ohishi, K., R. Aiyama, et al. (2001), Chem Pharm Bull (Tokyo) 49(7): 830-9.
Abstract: A novel series of acyl-CoA: cholesterol O-acyltransferase (ACAT) inhibitors were synthesized from a lead compound, 1-(4-hydroxy-3-methoxyphenyl)-7-phenylhept-1-en-3-one (1, Yakuchinone B) through a modification of three regions (A, B, C) in the molecule. In this study, the compounds prepared were tested for in vitro inhibitory activity on microsomal ACAT from the liver of rats and for in vivo hypocholesterolemic activity in rats given a high cholesterol diet. N-(3,5-Dimethoxy-4-n-octyloxycinnamoyl)-N'-(3,4-dimethylphenyl)piperazine (45), which belongs to the amide compounds, has finally been discovered. Compound 45 inhibited rat hepatic ACAT in a more striking manner than CI-976, an amide compound ACAT inhibitor, and it exhibited a high level of hypocholesterolemic activity in vivo. Since 45 strongly inhibited both microsomal ACAT prepared from HepG2 (a cell line derived from human hepatocarcinoma) and Caco2 (a cell line derived from human colon adenocarcinoma), there is speculation that 45 might have the ability to inhibit ACAT in both the human intestine and liver independent of the difference in the distribution of ACAT isozymes. On the other hand, 45 did not induce adrenotoxicity in subacute toxicity studies in rats. These results suggest that it has promise for development as a new therapeutic agent for hypercholesterolemia and atherosclerosis.

Structures of amphotericin B-cholesterol complex
Khutorsky, V. E. (1992), Biochim Biophys Acta 1108(2): 123-7.
Abstract: The structures of amphotericin B-cholesterol complex that forms a channel in a lipid membrane were analysed by molecular mechanics calculations. The symmetric complex consisting of eight rigid antibiotic and cholesterol molecules was considered. The presence of a continuous set of low-energy states of the complex with different values of the channel diameter was shown. These states are characterized by significant tilt of the amphotericin planes to the radial axis of the channel and by strong interaction between the charged ammonium and carboxyl groups of the antibiotic. Changes of the channel diameter may result in changes in pore permeability.

Structure-specific inhibition of lysosomal cholesterol transport in macrophages by various steroids
Aikawa, K., T. Furuchi, et al. (1994), Biochim Biophys Acta 1213(2): 127-34.
Abstract: Cultured mouse peritoneal macrophages effectively take up and metabolize liposomes containing phosphatidylserine and cholesterol, resulting in massive accumulation of cholesteryl esters and triacylglycerols in their cytoplasm (Nishikawa, K., Arai, H. and Inoue, K. (1990) J. Biol. Chem. 265, 5226-5231). With this system, various steroid derivatives were assessed as to their ability to inhibit the cholesteryl ester formation from endocytosed cholesterol in macrophages. Among the steroids tested, one group of steroids having an oxo group at the C17 or C20 position, such as androstenedione, dehydroisoandrosterone, progesterone and pregnenolone, completely inhibited cholesteryl ester formation at 10 microM. Another group of steroids having a hydroxy group at the C17 position, such as testosterone and androstenediol, had a lesser effect; complete inhibition of cholesteryl ester formation was achieved with 100 microM or more. The mechanism underlying the inhibition by the former class of steroids was further studied. These steroids did not block the uptake or lysosomal hydrolysis of liposomes, nor esterification of fatty acyl chains into triacylglycerols. Moreover, dehydroisoandrosterone and pregnenolone, both of which possess a hydroxy group at the C3 position, had essentially no effect on 25-hydroxycholesterol-stimulated esterification of endogenous cellular cholesterol. On the other hand, androstenedione and progesterone, which possess an oxo group at the C3 position, had a mild inhibitory effect on the esterification of endogenous cholesterol. Upon incubation with a steroid having an oxo group at the C17 or C20 position, free cholesterol taken up by macrophages was accumulated in phagolysosomes, as judged from cytochemical study with filipin-cholesterol staining. These results indicate that a series of structurally-related steroids characterized by the presence of an oxo group at the C17 or C20 position inhibit cholesteryl ester formation in macrophages through blocking the intracellular transport of endocytosed cholesterol from lysosomes to endoplasmic reticulum.

Student athlete cholesterol screening during routine precompetition examination
Kyle, J. M., R. B. Walker, et al. (1991), J Fam Pract 33(2): 172-6.
Abstract: BACKGROUND. When properly organized and conducted, the preseason physical examination process for scholastic athletes can identify existing medical and musculoskeletal problems as well as provide for age-specific anticipatory guidance. This examination may also present an ideal opportunity to screen for adolescent hyperlipidemia. METHODS. Seven-hundred seventy-seven (777) students, aged 11 to 15 years, from seven junior high schools received fingerstick cholesterol screening during a complete preseason physical examination. Elevated values were verified by repeat examination. Values were compared with previously published national norms for this age group. All students received information on cholesterol, and the parents and pediatrician or family physician of those with confirmed positive tests (higher than 4.8 mmol/L 185 mg/dL) were notified. RESULTS. One hundred fourteen (114), or approximately 15%, of the subjects were found to have elevated cholesterol levels. Of the 74 who returned for a second test, 38 (51%) were confirmed as having elevated cholesterol levels. Feedback from parents, principals, and coaches regarding the value of the screening and the associated education was overwhelmingly positive. CONCLUSIONS. In our experience, the precompetition examination provides an opportunity to screen for elevated cholesterol levels and to educate young people about hyperlipidemia.

Studies of alteration of hepatic cholesterol metabolism in puromycin-induced nephrotic syndrome in rats
Thabet, M. A., A. Challa, et al. (1993), Kidney Int 44(4): 789-94.
Abstract: Hypercholesterolemia frequently accompanies the nephrotic syndrome, but the mechanism responsible for elevation of plasma cholesterol is poorly understood. Specifically, the contribution of abnormal hepatic cholesterol metabolism to elevated concentrations of serum cholesterol has never been studied in depth. The objective of the present study was to define the alteration of hepatic cholesterol metabolism in puromycin induced nephrotic syndrome in rats. Studies involved measurements of specific activities of four enzymes participating in the maintenance of hepatic cholesterol metabolism: HMG-CoA-reductase, the rate limiting enzyme of cholesterol synthesis; cholesterol 7 alpha-hydroxylase, the rate limiting enzyme in bile acid synthesis; acyl CoA: cholesterol acyltransferase, the enzyme responsible for esterification of cholesterol; and cholesterol ester hydrolase (CEH), an enzyme which hydrolyzes cholesterol. Multiple injections of puromycin resulted in a production of nephrotic syndrome with massive proteinuria, hypoalbuminemia, hypercholesterolemia, ascites and edema. HMG-CoA-reductase (nmol/hr/mg protein) and cholesterol 7 alpha-hydroxylase activities (nmol/hr/mg protein) in rats with nephrotic syndrome were not statistically significant as compared to control rats (4.0 +/- 0.7 and 2.0 +/- 0.6 vs. 3.3 +/- 0.4 and 1.6 +/- 0.2), respectively. Our results also demonstrate, for the first time, that the normal diurnal rhythm in HMG-CoA reductase activity is no longer present in the nephrotic animals. The activities in the nephrotics in the day was 4.0 nmol/hr/mg and at night, 3.9 nmol/hr/day, compared to the control values of 3.3 nmol/hr/mg in the day and 6.9 nmol/hr/mg at night. ACAT activities were 428 +/- 78 versus 302 +/- 64 pmol/min/mg/protein (P = NS).(ABSTRACT TRUNCATED AT 250 WORDS)

Studies of cholesterol and bile acid metabolism, and early atherogenesis in hamsters fed GT16-239, a novel bile acid sequestrant (BAS)
Wilson, T. A., R. J. Nicolosi, et al. (1998), Atherosclerosis 140(2): 315-24.
Abstract: The purpose of this study was to compare the efficacy of GT16-239, an alkylated, cross-linked poly(allylamine) bile acid sequestrant with cholestyramine on cholesterol and bile acid metabolism, and early aortic atherosclerosis in hypercholesterolemic male F1B Golden Syrian hamsters. In this controlled study, 42 hamsters were divided into six groups and were fed a chow-based hypercholesterolemic diet supplemented with a 10% oil blend (55% coconut/45% corn), 0.1% cholesterol (w/w) (control) and either 0.9 or 1.2% cholestyramine or 0.2, 0.4 or 0.6% GT16-239 for 13 weeks. Laboratory analyses included evaluating plasma lipoprotein cholesterol and triglyceride concentrations, hepatic HMG-CoA reductase and 7 alpha-hydroxylase activities, fecal excretion of bile acids and neutral sterols, hepatic cholesterol concentrations, and early atherosclerosis (aortic fatty streak area). Relative to the control diet, the 0.6% GT16-239 versus the 1.2% cholestyramine significantly inhibited the elevation of plasma lipoprotein total cholesterol (TC) (-69% vs -40%), high density lipoprotein-cholesterol (HDL-C) (-49% vs -30%), and non-HDL-C (-81 vs -48%) concentrations; increased the activities of both HMG-CoA reductase (1492% vs 62%) and 7 alpha-hydroxylase (175% vs 86%); lowered the concentration of hepatic cholesteryl ester (-94% vs -59%); increased fecal cholesterol concentration (+28% vs -10%); and decreased aortic fatty streak area (-100% vs -86%). Unexpected findings of this comparison were increased fecal concentrations of cholic acid (533%) and chenodeoxycholic acid (400%) and the reduction in lithocholic acid (-50%) in the 0.6% GT16-239 compared to the 1.2% cholestyramine group. In summary, GT16-239 had a greater impact on cholesterol metabolism and early atherosclerosis in hypercholesterolemic hamsters than cholestyramine.

Studies of total and high density lipoprotein cholesterol in childhood malaria: a preliminary study
Agbedana, E. O., L. S. Salimonu, et al. (1990), Ann Trop Med Parasitol 84(5): 529-30.

Studies on cholesterol accumulation in radicular cyst fluid--origin of heat-stable cholesterol-binding protein
Yashima, M., N. Ogura, et al. (1990), Int J Biochem 22(2): 165-9.
Abstract: 1. The amount of apolipoprotein B (apo B) was measured using slit-immunoblotting in 20 specimens of radicular cyst fluids. Apo B was detected in all the cyst fluids with varying amounts. 2. Relationship between the amounts of apo B and free cholesterol or activity of heat-stable cholesterol-binding protein (HCBP) were examined. The amount of apo B was correlated well with the activity of HCBP (n = 20, r = 0.72, P less than 0.01) and with the amount of free cholesterol (n = 20, r = 0.45, P less than 0.05). 3. Anti-human apo B antibody inhibited cholesterol-binding activity in radicular cyst fluid. 4. When human-serum was chromatographed on a HPLC ion-exchange column, both cholesterol-binding activity and apo B had exactly the same retention time. 5. These results suggest that HCBP originates from beta-lipoprotein, and beta-lipoprotein may have an important role in cholesterol accumulation on radicular cysts.

Studies on enzymes involved in cholesterol metabolism using inhibitors
Tomoda, H. (1994), Tanpakushitsu Kakusan Koso 39(9): 1532-47.

Studies on serum levels of normal and cholesterol-fed Beijing duck apo A-I and apo B
Wu, X. and K. Wang (1991), Zhongguo Yi Xue Ke Xue Yuan Xue Bao 13(4): 246-50.
Abstract: Rabbit anti-Beijing duck apo A-I and apo B antisera were prepared by immunizing New Zealand rabbits with duck apo A-I and LDL, respectively. Rocket immunoelectrophoresis and radial immunodiffusion were used to determine the serum levels of apo A-I and apo B of 55 normal Beijing ducks and 8 cholesterol-fed Beijing ducks. Normal ducks contained about 145 mg apo A-I and 73 mg apo B per 100 ml serum; cholesterol-fed ducks contained about 167 mg apo A-I and 75 mg apo B per 100 ml serum.

Studies on the biosynthesis of polyisoprenols, cholesterol and ubiquinone in highly differentiated human hepatomas
Eggens, I., T. J. Ekstrom, et al. (1990), J Exp Pathol (Oxford) 71(2): 219-32.
Abstract: Surgical samples of human hepatic tissue were analysed morphologically and biochemically and highly differentiated hepatomas were compared with two control groups: morphologically normal liver tissue surrounding the tumour, and tissue from normal livers. In tumour homogenates cholesterol levels were more than twice, ubiquinone levels about half and the concentration of free dolichol about 10% of the control value. The levels of dolichyl phosphate were basically similar, whereas the phospholipid level was slightly lower in the tumours. In microsomes isolated from hepatomas, the level of cholesterol was about 30% higher than the control value. HMG-CoA reductase activity in microsomes isolated from hepatomas was elevated almost 100% in comparison to control. In hepatomas, no major alterations in the compositions of dolichol or dolichyl phosphate could be observed. The relative amounts of alpha-saturated and alpha-unsaturated polyprenols were also basically unaltered in hepatomas. Liver samples were incubated with 3H-mevalonic acid and radioactivity was monitored in polyprenols. With control tissue, incorporation was considerably higher in alpha-unsaturated polyprenols than in their alpha-saturated counterparts. In the tumours the rates of incorporation into both polyprenol fractions were much lower, although still higher in the alpha-unsaturated fraction. Labelling of polyisoprenols containing 19 isoprene residues was higher than that of 20 residues. The pattern of labelling in the polyisoprenyl-P fraction was similar. In hepatomas the incorporation into cholesterol and ubiquinone-10 was about 100% higher and 50% lower respectively compared with control tissue. The results in this study of hepatomas indicate that the levels of various lipids may be influenced not only by the regulatory enzyme HMG-CoA reductase, but also by other enzymes catalysing reactions subsequent to this regulatory point. It is also suggested that levels of cholesterol, ubiquinone and dolichol may be regulated independently subsequent to the branch point at farnesylpyrophosphate.

Studies on the effect of feeding cupric sulfate pentahydrate to laying hens on egg cholesterol content
Pesti, G. M. and R. I. Bakalli (1998), Poult Sci 77(10): 1540-5.
Abstract: Two experiments were conducted to test the hypothesis that pharmacological levels of dietary Cu could reduce egg cholesterol content. White Leghorn hens 30 to 39 wk of age were fed corn and soybean meal diets with 0, 125, or 250 mg supplemental Cu/kg diet from cupric sulfate pentahydrate (basal diet = 6.74 mg Cu/kg). Body weight, feed consumption, egg weights, egg specific gravity, and Haugh Units were not consistently affected during the 8-wk feeding trials. Egg production was significantly increased (P < 0.05) in the second 4-wk period by supplemental Cu in both experiments. Egg yolk cholesterol concentrations were decreased by feeding 125 mg Cu/kg diet (11.7 vs 8.6 mg/g, average of two experiments); feeding 250 mg Cu/kg resulted in further declines in egg cholesterol but the differences were not significant (7.9 mg/g). Changes in plasma cholesterol concentrations were similar to those of yolk cholesterol. Small but significant amounts of Cu accumulated in the yolks and shells of eggs from Cu-supplemented hens; however, most of the Cu fed was found in the excreta.

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Cholesterol Journal Articles