Abstract：
:Inositol lipid kinases have been studied extensively in both plant and animal systems. However, major limitations for in vitro studies of recombinant lipid kinases are the low specific activity and instability of the purified proteins. Our goal was to determine if cyclodextrins would provide an effective substrate delivery system and enhance the specific activity of lipid kinases. For these studies, we have used recombinant Arabidopsis thaliana phosphatidylinositol phosphate kinase 1 (At PIPK1). At PIPK1 was produced as a fusion protein with glutathione-S-transferase and purified on glutathione-Sepharose beads. A comparison of lipid kinase activity using substrate prepared in alpha-, beta-, or gamma-cyclodextrin indicated that beta-cyclodextrin was most effective and enhanced lipid kinase activity 6-fold compared with substrate prepared in Triton X-100-mixed micelles. We have optimized reaction conditions and shown that product can be recovered from the cyclodextrin-treated recombinant protein, which reveals a potential method for automating the assay for pharmacological screening.
journal_name
J Lipid Resjournal_title
Journal of lipid researchauthors
Davis AJ,Perera IY,Boss WFdoi
10.1194/jlr.D400005-JLR200keywords:
subject
Has Abstractpub_date
2004-09-01 00:00:00pages
1783-9issue
9eissn
0022-2275issn
1539-7262pii
D400005-JLR200journal_volume
45pub_type
杂志文章abstract：:Cerebrotendinous xanthomatosis (CTX) is a treatable neurodegenerative metabolic disorder of bile acid synthesis in which symptoms can be prevented if treatment with chenodeoxycholic acid supplementation is initiated early in life, making CTX an excellent candidate for newborn screening. We developed a new dried blood ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.P075051
更新日期：2017-05-01 00:00:00
abstract：:A simple procedure is described for preparing lyso-GM1, a GM1 derivative that lacks the fatty acid moiety, starting from GM1 ganglioside using a one-pot reaction. Ganglioside deacylation was carried out in KOH/propan-1-ol in the absence of oxygen. The yield of lyso-GM1 under optimal conditions (6 h, 90 degrees C, 0.2 ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1992-08-01 00:00:00
abstract：:Previous studies have shown that antibodies to cubulin, a receptor on the yolk sac that binds high density lipoproteins (HDL) and cobalamin, induce fetal abnormalities. Mice with markedly low concentrations of plasma HDL-cholesterol (HDL-C) give birth to healthy pups, however. To establish whether maternal HDL-C has a...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：2000-03-01 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：:Lipoprotein lipase (LPL) and hepatic lipase (HL) mediate the hydrolysis of triglycerides and phospholipids present in circulating lipoprotein particles and are essential for normal lipid metabolism. Both enzymes have a similar primary amino acid structure and share requirements for intact catalytic, lipid binding, and...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1993-08-01 00:00:00
abstract：:A modification of the semiautomated assay method of Antonis (1965. J. Lipid Res. 6:307-312) for free fatty acid is presented. Free fatty acids are extracted from serum or plasma into di-n-butyl ether-2-methoxyethanol; the extract is almost free from phospholipids. The acids are analyzed in a portion of extract by a co...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1978-02-01 00:00:00
abstract：:Inhibition of Ehrlich ascites carcinoma respiration by aqueous extracts of oxidized linoleic or linolenic acid (aqueous emulsions UV-irradiated, 90 min) was associated entirely with relatively involatile compounds which were both thiobarbituric acid (TBA)-reactive and peroxidase-reactive. Inhibitory compounds were hea...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1966-05-01 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：:Treatment of native human Lp(a) under nondenaturing conditions with dithiothreitol yielded both a lipoprotein particle and a lipid-free protein component that could be separated by either ultracentrifugation at d 1.063 g/ml or heparin-Sepharose chromatography. The protein component only showed antigenicity against ant...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1985-11-01 00:00:00
abstract：:This review presents an overview of mammalian phospholipid synthesis and the cellular locations of the biochemical activities that produce membrane lipid molecular species. The generalized endoplasmic reticulum compartment is a central site for membrane lipid biogenesis, and examples of the emerging relationships betw...
journal_title：Journal of lipid research
pub_type： 杂志文章,评审
doi：10.1194/jlr.R800049-JLR200
更新日期：2009-04-01 00:00:00
abstract：:Autosomal dominant Stargardt-like macular dystrophy (STGD3) in humans results from mutations in elongation of very long chain FAs-like 4 (ELOVL4), which leads to vision loss in young adults. ELOVL4 is an integral endoplasmic reticulum (ER) protein that mediates the elongation of very long chain (VLC) FAs. Mutations in...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M045443
更新日期：2014-04-01 00:00:00
abstract：:Hormone-sensitive lipase (HSL) catalyzes the rate-limiting step in adipocyte lipolysis. The activity of HSL is thought to be primarily regulated by reversible phosphorylation. However, the regulation of HSL activity by pre-translational mechanisms has been poorly studied. The present studies were undertaken to explore...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1998-08-01 00:00:00
abstract：:N-3 trans geometrical isomers of 20:5 n-3 and 22:6 n-3 were isolated from rats fed heated linseed oil. The ability of these acids to inhibit 20:4 n-6 metabolism by human platelets was examined. The concentrations required to inhibit 50% of platelet aggregation (IC50) induced by 2.5 microM 20:4 n-6 were higher for the ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1990-07-01 00:00:00
abstract：:Increased plasma cholesterol is a known risk factor for cardiovascular disease. Lipoprotein particles transport both cholesterol and triglycerides through the blood. It is thought that the size distribution of these particles codetermines cardiovascular disease risk. New types of measurements can determine the concent...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M800354-JLR200
更新日期：2009-12-01 00:00:00
abstract：:The action of cycloheximide on some parameters of glucose and lipid metabolism was studied in vitro in epidiymal fat pads from fasted rats. Incubation of fat pads with cycloheximide (1 mug/ml) for 2 hours resulted in a two-fold increase in glucose uptake, glucose oxidation, incorporation of glucose into lipids, and re...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1977-01-01 00:00:00
abstract：:Some commercial preparations of common natural conjugated bile salts contain impurities (e.g., amines, lipids, and calcium) that are likely to affect their physicochemical properties. A method was developed for purifying commercial preparations of sodium salts of glycine- and taurine-conjugated bile acids. The method ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1995-12-01 00:00:00
abstract：:Synthesis of apolipoprotein (apo)E in hepatocytes leads to both secretion and retention in cell surface pools. Inclusion of Brefeldin A to HepG2 cells prompted a rapid decrease of cell surface apoE to about 37% of control values after a 3-h incubation. The t(1/2) for this dynamic pool was estimated to be 15 min. In co...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：2001-09-01 00:00:00
abstract：:Acylglycerophosphate acyltransferase 4 (AGPAT4)/lysophosphatidic acid acyltransferase delta catalyzes the formation of phosphatidic acid (PA), a precursor of triacyl-glycerol (TAG). We investigated the effect of Agpat4 gene ablation on white adipose tissue (WAT) after finding consistent expression across depots. Epidi...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M079152
更新日期：2017-10-01 00:00:00
abstract：:Our previous studies have demonstrated the activity of oncostatin M (OM) in stimulating the transcription of the human LDL receptor (LDLR) gene in HepG2 cells through a sterol-independent regulatory mechanism. The current studies were designed to determine whether this in vitro property of OM could be recapitulated in...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M400425-JLR200
更新日期：2005-06-01 00:00:00
abstract：:The diurnal cycle of hepatic HMG CoA reductase activity was studied under conditions of controlled feeding where the percentage of dietary lipid, alone or in combination with 2% cholestyramine, was varied. Cholestyramine caused an increase in HMG CoA reductase activity that began soon after feeding started and peaked ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1972-11-01 00:00:00
abstract：:Apolipoprotein A-V (apoA-V) first gained attention as a regulator of triglycerides through transgenic mouse studies. Furthermore, peroxisome proliferator-activated receptor alpha (PPARalpha) agonists such as fenofibrate increase apoA-V mRNA expression. Our group recently developed the first assay to quantitate serum a...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.C500010-JLR200
更新日期：2005-08-01 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：:When grown in the presence of bile acids, two strains of Clostridium limosum were found to contain significant amounts of NADP-dependent 7 alpha/7 beta-hydroxysteroid dehydrogenase and NAD-dependent 7 alpha-hydroxysteroid dehydrogenase which were active against conjugated and unconjugated bile acids. No measurable act...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1985-03-01 00:00:00
abstract：:The total nitrogen and L-hydroxyproline concentrations of two intra-abdominal and two subcutaneous sites of adipose tissue were measured in male and female Wistar rats. At 100 g body weight the nitrogen concentration of intra-abdominal adipose tissue was slightly lower than that of subcutaneous adipose tissue. As age ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1970-07-01 00:00:00
abstract：:Infusions of 2-deoxyglucose (2-DG) into intact, adrenalectomized, and adrenalectomized-hypophysectomized dogs caused increases in plasma free fatty acid (FFA) levels which could be reversed by infusing hexamethonium, or prevented by epidural anesthesia or destruction of the thoracic spinal cord. Similar infusions of 2...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1966-05-01 00:00:00
abstract：:The behavior of human omental fat cells in vitro has been examined in order to define conditions under which glucose is converted to glyceride-glycerol and glyceride fatty acids. Synthesis of glyceride fatty acids from glucose reached maximal rates only after several hours of incubation in Krebs-Henseleit bicarbonate ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1969-05-01 00:00:00
abstract：:Previous in vitro studies have revealed that oxidized low density lipoprotein (OxLDL) has negative effects on the proliferation and activity of endothelial progenitor cells (EPCs). Here, we evaluated the effect of OxLDL on the therapeutic potential of EPCs in ischemia-induced neovascularization. EPCs derived from mobi...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M600251-JLR200
更新日期：2007-03-01 00:00:00
abstract：:Triglyceride (TG) concentration is used as a marker of cardiometabolic risk. However, diurnal and possibly weekday variation exists in TG concentrations. The objective of this work was to investigate weekday variation in TG concentrations among 1.8 million blood samples drawn between 2008 and 2015 from patients in the...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M074062
更新日期：2017-06-01 00:00:00
abstract：:Apolipoprotein A-II (apoA-II) is the second major apolipoprotein following apolipoprotein A-I (apoA-I) in HDL. ApoA-II has multiple physiological functions and can form senile amyloid fibrils (AApoAII) in mice. Most circulating apoA-II is present in lipoprotein A-I/A-II. To study the influence of apoA-I on apoA-II and...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M013235
更新日期：2011-08-01 00:00:00
abstract：:Leukotriene B4 (LTB4) is a potent lipid mediator of inflammation that acts primarily via a seven-transmembrane-spanning, G-protein-coupled receptor denoted BLT1. Here, we describe the synthesis and characterization of fluorescent analogs of LTB4 that are easy to produce, inexpensive, and without the disadvantages of a...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.D500005-JLR200
更新日期：2005-06-01 00:00:00
