Abstract：
:Sphingolipids comprise a highly diverse and complex class of molecules that serve not only as structural components of membranes but also as signaling molecules. To understand the differential role of sphingolipids in a regulatory network, it is important to use specific and quantitative methods. We developed a novel LC-MS/MS method for the rapid, simultaneous quantification of sphingolipid metabolites, including sphingosine, sphinganine, phyto-sphingosine, di- and trimethyl-sphingosine, sphingosylphosphorylcholine, hexosylceramide, lactosylceramide, ceramide-1-phosphate, and dihydroceramide-1-phosphate. Appropriate internal standards (ISs) were added prior to lipid extraction. In contrast to most published methods based on reversed phase chromatography, we used hydrophilic interaction liquid chromatography and achieved good peak shapes, a short analysis time of 4.5 min, and, most importantly, coelution of analytes and their respective ISs. To avoid an overestimation of species concentrations, peak areas were corrected regarding isotopic overlap where necessary. Quantification was achieved by standard addition of naturally occurring sphingolipid species to the sample matrix. The method showed excellent precision, accuracy, detection limits, and robustness. As an example, sphingolipid species were quantified in fibroblasts treated with myriocin or sphingosine-kinase inhibitor. In summary, this method represents a valuable tool to evaluate the role of sphingolipids in the regulation of cell functions.
journal_name
J Lipid Resjournal_title
Journal of lipid researchauthors
Scherer M,Leuthäuser-Jaschinski K,Ecker J,Schmitz G,Liebisch Gdoi
10.1194/jlr.D005322subject
Has Abstractpub_date
2010-07-01 00:00:00pages
2001-11issue
7eissn
0022-2275issn
1539-7262pii
jlr.D005322journal_volume
51pub_type
杂志文章abstract：:HDLs have been proposed to have antiatherogenic properties because of their role in reverse cholesterol transport as lipid acceptors. To elucidate the phospholipid profile of these particles, we used electrospray ionization mass spectrometry to examine the phosphatidylcholine (PC) and sphingomyelin (SM) composition of...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M500011-JLR200
更新日期：2005-06-01 00:00:00
abstract：:Animal studies have suggested that angiopoietin-like 4 (Angptl4) regulates adiposity through central and peripheral mechanisms. The aim of this study was to investigate whether serum concentration and adipose tissue expression of Angptl4 are associated with obesity-related parameters in humans. Altogether, 75 dizygoti...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.P015867
更新日期：2011-08-01 00:00:00
abstract：:Fungal lipases and human pancreatic lipase (hPL) share a common tertiary structure termed the alpha/beta hydrolase fold. In contrast, the region C-terminal to the common tertiary structure does not share any common structural features with fungal lipases, leading to the hypothesis that the divergent C-terminal domain ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1995-05-01 00:00:00
abstract：:Monolayers of Caco-2 cells, a human enterocyte cell line, were incubated with [1-14C]15-hydroxyeicosatetraenoic acid (15-HETE), a lipid mediator of inflammation, and [1-14C]arachidonic acid. Both fatty acids were taken up readily and metabolized by Caco-2 cells. [1-14C]Arachidonic acid was directly esterified in cellu...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1990-05-01 00:00:00
abstract：:The lipids of rat milk, the contents of 9-- 10-day-old rat stomach and intestine, lymph, plasma, and liver were quantitated and their fatty acids were analyzed. Rat milk consists of 97% triacylglycerols, of which 35% of the fatty acids are of medium chain length (C8-C12). However, stomach triacylglycerols show a 25% r...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1981-05-01 00:00:00
abstract：:A preparation of cerebral microvessels was used to demonstrate the presence of lipoprotein lipase and acid lipase activity in the microvasculature of rabbit brain. Microvessels, consisting predominantly of capillaries, small arterioles, and venules, were islated from rabbit brain. Homogenates were assayed for lipolyti...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1979-05-01 00:00:00
abstract：:Cystic fibrosis (CF) patients and model systems exhibit consistent abnormalities in PUFA metabolism, including increased metabolism of linoleate to arachidonate. Recent studies have connected these abnormalities to increased expression and activity of the Δ6- and Δ5-desaturase enzymes. However, the mechanism connectin...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M050369
更新日期：2014-07-01 00:00:00
abstract：:Previous studies have reported both positive and negative effects of culture of islets at high glucose concentrations on regulated insulin secretion. Here, we have reexamined this question in mouse islets and determined the role of changes in lipid synthesis in the effects of glucose. Glucose-stimulated insulin secret...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M700533-JLR200
更新日期：2008-04-01 00:00:00
abstract：:Recent discovery and characterization of APOAV suggests a role in metabolism of triglyceride (TG)-rich lipoproteins. Previously, variation at the APOAV locus was shown to modestly influence plasma TGs in normolipidemic samples. The aims of this study were to assess the effects of a polymorphism in APOAV (T-1131C) in t...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M200480-JLR200
更新日期：2003-06-01 00:00:00
abstract：:Distinct isoforms of long-chain acyl-CoA synthetases (ACSLs) may partition fatty acids toward specific metabolic cellular pathways. For each of the five members of the rat ACSL family, we analyzed tissue mRNA distributions, and we correlated the mRNA, protein, and activity of ACSL1 and ACSL4 after fasting and refeedin...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M600150-JLR200
更新日期：2006-09-01 00:00:00
abstract：:We report the molecular basis of familial chylomicronemia and recurrent pancreatitis in five members of a large Dutch family. All patients had normal plasma hepatic lipase and apoC-II levels, but absent lipoprotein lipase (LPL) catalytic activity and low LPL mass in postheparin plasma. The mutation in the LPL gene was...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1993-12-01 00:00:00
abstract：:We investigated, in the present study, the role of reactive oxygen intermediates (ROI) in the control of macrophage lipoprotein lipase (LPL) secretion. Exposure of murine macrophages to increasing concentrations of hydrogen peroxide (H2O2) resulted in enhanced basal LPL production and mRNA levels. The increase of LPL ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1996-04-01 00:00:00
abstract：:Proprotein convertase subtilisin/kexin type 9 (PCSK9) is a secreted protein that regulates hepatic low-density lipoprotein receptor (LDLR) levels in humans. PCSK9 has also been shown to regulate the levels of additional membrane-bound proteins in vitro, including the very low-density lipoprotein receptor (VLDLR), apol...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M006635
更新日期：2010-09-01 00:00:00
abstract：:The novel acyl-CoA:cholesterol acyltransferase (ACAT) inhibitor CP-113,818 has been characterized in vitro against ACAT isolated from liver and intestine from a variety of species including human subjects, and in vivo in cholesterol-fed rats, hamsters, rabbits, and two species of nonhuman primates. CP-113,818 is a pot...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-10-01 00:00:00
abstract：:Gap junctions of rat brown adipose tissue (BAT) were studied with the freeze-fracture technique during development. The frequency and the mean area of gap junctions increased after birth, reached a maximal development during the functional phase of the tissue (birth to 3--4 weeks) and decreased while brown adipose tis...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1980-07-01 00:00:00
abstract：:Abetalipoproteinemia (ABL) and chylomicron retention disease (CMRD) are extremely rare recessive forms of hypobetalipoproteinemia characterized by intestinal lipid malabsorption and severe vitamin E deficiency. Vitamin E is often supplemented in the form of fat-soluble vitamin E acetate, but fat malabsorption consider...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M085043
更新日期：2018-09-01 00:00:00
abstract：:Hyperlipidemia is a risk factor for development and progression of diabetic nephropathy. However, it is not known if reduction of hyperlipidemia is protective against progression of disease. The goal of this study was to determine if reduction of hypercholesterolemia could limit progression of diabetic nephropathy. Di...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M002972
更新日期：2010-06-01 00:00:00
abstract：:Adipose tissue development was studied histochemically and histologically in fetuses from lean and obese sows. At 110 days of gestation, fetuses were removed from Ossabaw (obese-feral) sows and from sows selected for high backfat (obese-domestic) and for low backfat (lean) thickness. Body weights were similar for lean...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1983-03-01 00:00:00
abstract：:Human striated muscle samples, from male control and Duchenne muscular dystrophy-affected children, were subjected to cluster-time-of-flight secondary ion mass spectrometry (cluster-ToF-SIMS) imaging using a 25 keV Bi(3)(+) liquid metal ion gun under static SIMS conditions. Spectra and ion density maps, or secondary i...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M700421-JLR200
更新日期：2008-02-01 00:00:00
abstract：:The characteristics of 7 alpha-dehydroxylase, a bile acid-biotransforming enzyme, were determined using dialyzed cell extracts of Eubacterium sp. V.P.I. 12708. 7 alpha-Dehydroxylase was induced by cholic acid in this organism. Induction by cholic acid resulted in the differential synthesis of at least five new polypep...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1981-08-01 00:00:00
abstract：:The relative rates of hydrolysis of the secondary ester in glycerol 1,3-benzylidene 2-oleate and in glycerol 1,3-dihexadecyl ether 2-oleate, and of the primary and secondary esters in triolein were determined. Both unaltered and selectively inactivated rat pancreatic juice were used as sources of enzyme. It was found ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1968-01-01 00:00:00
abstract：:Human lysosomal acid lipase/cholesteryl ester hydrolase (hLAL) is essential for the intralysosomal metabolism of cholesteryl esters and triglycerides taken up by receptor-mediated endocytosis of lipoprotein particles. The key role of the enzyme in intracellular lipid homeostasis is illustrated by two lysosomal storage...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1999-02-01 00:00:00
abstract：:We have studied the synthesis, modification, and secretion of rat apoE in primary cultures of neonatal aortic smooth muscle cells and adult rat hepatocytes. The cultures were pulsed with [35S]methionine and the intracellular and secreted apoE were immunoprecipitated and analyzed by two-dimensional isoelectric focusing...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1988-07-01 00:00:00
abstract：:A reversed phase high performance liquid chromatographic (HPLC) method was developed for direct profiling and determination of adrenal cholesteryl ester composition. Cholesteryl adrenate and cholesteryl cervonate, which are not commercially available, were synthesized as markers. Lipid extracts of rat adrenal homogena...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-06-01 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：:Adipocyte differentiation and function have become areas of intense focus in the field of energy metabolism; however, understanding the role of specific genes in the establishment and maintenance of fat cell function can be challenging and complex. In this review, we offer practical guidelines for the study of adipocy...
journal_title：Journal of lipid research
pub_type： 杂志文章,评审
doi：10.1194/jlr.R046441
更新日期：2014-04-01 00:00:00
abstract：:ATP-binding cassette transporter A1 (ABCA1) plays a crucial role in apoA-I lipidation, a key step in reverse cholesterol transport. cAMP induces apoA-I binding activity and promotes cellular cholesterol efflux. We investigated the role of the cAMP/protein kinase A (PKA) dependent pathway in the regulation of cellular ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.m200235-jlr200
更新日期：2002-12-01 00:00:00
abstract：:The fatty acid transport protein (FATP) Fat1p in the yeast Saccharomyces cerevisiae functions in concert with acyl-coenzyme A synthetase (ACSL; either Faa1p or Faa4p) in vectorial acylation, which couples the transport of exogenous fatty acids with activation to CoA thioesters. To further define the role of Fat1p in t...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M700300-JLR200
更新日期：2007-11-01 00:00:00
abstract：:The apolipoprotein (apo) B-100-specific monoclonal antibody MB47 has been widely used in lipoprotein metabolism and atherosclerosis research. When bound to apoB-100 on low density lipoproteins (LDL), antibody MB47 completely blocks the binding of LDL to the LDL receptor. The epitope for antibody MB47 has previously be...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-03-01 00:00:00
abstract：:Lipid concentrations are heritable traits. Recently, the number of known genetic loci associated with lipid levels in adults increased from 95 to 157. The effects of these 157 loci have not been tested in children. Considering that lipid levels track from childhood to adulthood, we studied to determine whether these v...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.P066902
更新日期：2016-12-01 00:00:00
