Abstract：
:Membrane-protein interaction plays key roles in a wide variety of biological processes. Although various methods have been employed to measure membrane binding of soluble proteins, a robust high-throughput assay that is universally applicable to all proteins is lacking at present. Here we report a new fluorescence quenching assay utilizing enhanced green fluorescence protein (EGFP)-fusion proteins and a lipid containing a dark quencher, N-dimethylaminoazobenzenesulfonyl-phosphatidylethanolamine (dabsyl-PE). The EGFP fluorescence emission intensity showed a large decrease (i.e., >50%) when EGFP-fusion proteins bound the vesicles containing 5 mol% dabsyl-PE. This simple assay, which can be performed using either a cuvette-based spectrofluorometer or a fluorescence plate reader, allowed rapid, sensitive, and accurate determination of lipid specificity and affinity for various lipid binding domains, including two pleckstrin homology domains, an epsin N-terminal homology domain, and a phox homology domain. The assay can also be applied to high-throughput screening of small molecules that modulate membrane binding of proteins.
journal_name
J Lipid Resjournal_title
Journal of lipid researchauthors
Kim H,Afsari HS,Cho Wdoi
10.1194/jlr.D041376subject
Has Abstractpub_date
2013-12-01 00:00:00pages
3531-8issue
12eissn
0022-2275issn
1539-7262pii
jlr.D041376journal_volume
54pub_type
杂志文章abstract：:ZR-78 and ZR-82 cells are two peroxisomal-deficient Chinese hamster ovary (CHO) cell mutants. These cells lack normal peroxisomes and show reduced levels of plasmalogen synthesis and other peroxisomal functions attributed to the deficiency of peroxisomal matrix enzymes. As we have recently identified two HMG-CoA reduc...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1998-09-01 00:00:00
abstract：:The effect of inhibiting lysosomal protein degradation on the activity of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase was determined using a mouse mammary cell line (TS-85) which expresses a temperature-sensitive mutation in the ubiquitin degradative pathway. Incubating cells for 18 hr in medium containi...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1986-03-01 00:00:00
abstract：:Very few studies have investigated the interrelations between proprotein convertase subtilisin/kexin type 9 (PCSK9) metabolism, cholesterol synthesis, and cholesterol absorption. We aimed to address this issue in a large clinical trial of 245 patients with hypercholesterolemia. Serum lipids, PCSK9, lathosterol (choles...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.P088583
更新日期：2019-01-01 00:00:00
abstract：:Measurements of enzymatic activity have demonstrated that lipoprotein lipase (LPL), the principal enzyme responsible for hydrolysis of circulating triglyceride, is present in a number of tissues including brain, kidney, and adrenal gland. To determine the sites of synthesis of LPL in these tissues, in situ hybridizati...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1989-10-01 00:00:00
abstract：:Apolipoprotein E plays a critical role in plasma lipoprotein clearance. Peptide models of a highly conserved, N-terminal domain of this protein have been shown to increase the binding of low density lipoprotein (LDL) to fibroblast cell surfaces independently of the low density lipoprotein receptor. Here we provide dat...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1999-04-01 00:00:00
abstract：:The inverse relationship between plasma levels of high density lipoprotein (HDL) and coronary heart disease rates has suggested that HDL might influence body stores of cholesterol. Therefore, we have investigated potential relationships between the parameters of body cholesterol metabolism and the plasma levels of HDL...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1985-09-01 00:00:00
abstract：:The beta-oxidation of stearic acid and of alpha- and gamma-methyl isoprenoid-derived fatty acids (pristanic and tetramethylheptadecanoic acids, respectively) was investigated in normal skin fibroblasts and in fibroblasts from patients with inherited defects in peroxisomal biogenesis. Stearic acid beta-oxidation by nor...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1990-02-01 00:00:00
abstract：:Fatty acid elongases and desaturases play an important role in hepatic and whole body lipid composition. We examined the role that key transcription factors played in the control of hepatic elongase and desaturase expression. Studies with peroxisome proliferator-activated receptor alpha (PPARalpha)-deficient mice esta...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M600177-JLR200
更新日期：2006-09-01 00:00:00
abstract：:Spectrophotometric techniques for determining the activities of lipases, lysophospholipases, and phospholipases are reviewed. These methods involve the use of thioester substrate analogs as well as omega-nitrophenyl derivatives of the corresponding lipids. The most promising results are obtained with the thioester sub...
journal_title：Journal of lipid research
pub_type： 杂志文章,评审
doi：
更新日期：1984-12-15 00:00:00
abstract：:Previous studies from our laboratory have shown that oral administration of stigmastanyl-phosphocholine (Ro 16-6532) reduces plasma cholesterol levels in experimental animals on diets free of added cholesterol. In the present study, effects of Ro 16-6532 and lovastatin on lipoprotein levels and metabolism were investi...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1995-07-01 00:00:00
abstract：:When stimulated, rat serosal mast cells degranulate and secrete a cytoplasmic neutral protease, chymase. We studied the fragmentation of apolipoprotein (apo) A-I during proteolysis of HDL(3) by chymase, and examined how chymase-dependent proteolysis interfered with the binding of eight murine monoclonal antibodies (Ma...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：2000-06-01 00:00:00
abstract：:Conjugated linoleic acid (CLA), a mixture of isomers of linoleic acid, has previously been shown to be able to decrease porcine subcutaneous (SC) adipose tissue levels while increasing the count of intramuscular (IM) adipose tissue in vivo. However, the underlying mechanisms through which it acts are poorly understood...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M600525-JLR200
更新日期：2007-08-01 00:00:00
abstract：:An online, two-dimensional (2D) liquid chromatography (LC) quadrupole time-of-flight mass spectrometry (QToF-MS) method was developed for lipid profiling of rat peritoneal surface layers, in which the lipid classes and species could be simultaneously separated in one injection with a significantly increased sensitivit...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.D007567
更新日期：2010-09-01 00:00:00
abstract：:Insulin-dependent diabetes mellitus (IDDM) is associated with elevated plasma triglyceride levels that normalize after insulin administration. The observation that overexpression of the apoC-III gene in transgenic mice can cause hypertriglyceridemia and other evidence implicating apoC-III in the regulation of triglyce...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-11-01 00:00:00
abstract：:The enzymic incorporation of choline-1,2-(14)C from CDP-choline-1,2-(14)C into phosphatidylcholine by spinach leaf preparations was characterized. The enzyme catalyzing the incorporation, choline phosphotransferase, had a pH optimum of about 8.0 and required either Mn(2+) or Mg(2+) as cofactor. The saturation concentr...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1971-07-01 00:00:00
abstract：:Bile acids play important roles in the regulation of lipid, glucose, and energy homeostasis. Recent studies suggest that glucose regulates gene transcription in the liver. The aim of this study was to investigate the potential role of glucose in regulation of bile acid synthesis in human hepatocytes. High glucose stim...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M002782
更新日期：2010-04-01 00:00:00
abstract：:The accurate quantitation of high density lipoproteins has recently assumed greater importance in view of studies suggesting their negative correlation with coronary heart disease. High density lipoproteins may be estimated by measuring cholesterol in the plasma fraction of d > 1.063 g/ml. A more practical approach is...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1978-01-01 00:00:00
abstract：:The mRNA level for cytosolic NADP-dependent isocitrate dehydrogenase (IDH1) increases 2.3-fold, and enzyme activity of NADP-isocitrate dehydrogenase (IDH) 63%, in sterol-deprived HepG2 cells. The mRNA levels of the NADP- and NAD-dependent mitochondrial enzymes show limited or lack of regulation under the same conditio...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M300285-JLR200
更新日期：2003-11-01 00:00:00
abstract：:Changes in chromatin structure regulate gene expression and genome maintenance. Molecules that bind to the nucleosome, the complex of DNA and histone proteins, are key modulators of chromatin structure. Previous work indicated that cholesterol, a ubiquitous cellular lipid, may bind to chromatin in vivo, suggesting a p...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M074997
更新日期：2017-05-01 00:00:00
abstract：:The genetic variability of apolipoprotein E (apoE) influences plasma lipoprotein levels, and allele frequencies differ between African Americans and Caucasians. As African Americans have higher lipoprotein [a] (Lp[a]) levels than Caucasians, we investigated the effects of the apoE gene on allele-specific apolipoprotei...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M600431-JLR200
更新日期：2007-03-01 00:00:00
abstract：:The concentration of cholesterol carried in low density lipoproteins (LDL-C) is primarily determined by the rate at which LDL-C is produced (Jt) and the rate at which the liver takes up this particle through receptor-dependent transport (Jm). The accumulation of specific dietary fatty acids in the liver profoundly alt...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-09-01 00:00:00
abstract：:Familial hypobetalipoproteinemia, a syndrome associated with low plasma cholesterol levels, can be caused by apoB gene mutations. We identified a healthy 42-year-old man whose total plasma cholesterol level was 80 mg/dl. His plasma very low density lipoprotein (VLDL) contained a unique truncated apoB species, apoB-83,...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1992-04-01 00:00:00
abstract：:Circulating concentrations of mevalonic acid (MVA) change in parallel with, and may be used as a marker of cholesterol biosynthesis. Plasma MVA levels have been quantified using a sensitive and specific capillary gas chromatography-electron capture mass spectrometric assay. The detection limit for MVA in plasma is 100...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1991-06-01 00:00:00
abstract：:In skin fibroblasts grown from four children with a homozygous form of type II hyperlipoproteinemia, the feedback control of sterol synthesis and the inhibitory effect on hydroxymethylglutaryl (HMG) CoA reductase activity by serum or low density lipoprotein were present, though diminished compared with the effects in ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1975-03-01 00:00:00
abstract：:The pregnane X receptor (PXR, also known as SXR) is a nuclear hormone receptor activated by xenobiotics as well as diverse sterols and their metabolites. PXR functions as a xenobiotic sensor to coordinately regulate xenobiotic metabolism via transcriptional regulation of xenobiotic-detoxifying enzymes and transporters...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M017376
更新日期：2011-09-01 00:00:00
abstract：:As microsomal triglyceride transfer protein (MTP) is required for the assembly and secretion of apoB-containing lipoproteins by intestinal epithelial cells, the characterization and regulation of MTP in human intestinal cells, CaCo-2, was studied. CaCo-2 cells express MTP mRNA of 4.2 kb and a 97 kDa subunit of MTP pro...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1997-01-01 00:00:00
abstract：:Hepatocytes express on their surfaces more than one class of receptors capable of mediating the internalization of lipoproteins. However, relatively little is known about the binding characteristics of hepatic receptors for various lipoproteins, about the regulation of the receptors, and about the consequences for int...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1983-12-01 00:00:00
abstract：:The metabolism of chenodeoxycholic acid by Pseudomonas sp. NCIB 10590 under strict anaerobic conditions was studied. A range of unsaturated acidic and neutral metabolites were isolated and identified. The major acidic product was chola-4,6-dien-3-one-24-oic acid whilst the major neutral product was androsta-4,6-dien-3...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1983-09-01 00:00:00
abstract：:Hepatic free cholesterol levels are influenced by cholesterol synthesis and ester formation, which, in turn, might regulate cholesterol secretion into bile and plasma. We manipulated the rates of hepatic cholesterol synthesis and esterification and measured biliary and very low density lipoprotein (VLDL) cholesterol s...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1992-11-01 00:00:00
abstract：:Triacylglycerols (TG) are the major storage form of energy in eukaryotic organisms and are synthesized primarily by acyl CoA:1,2-diacylglycerol acyltransferase (DGAT) enzymes. In vitro DGAT activity has previously been quantified by measuring the incorporation of either radiolabeled fatty acyl CoA or diacylglycerol (D...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.D016626
更新日期：2011-09-01 00:00:00
