Abstract：
:The demand to study the cellular localization of specific lipids has led to recent advances in lipid probes and microscopy. Alkyne lipids bear a small, noninterfering tag and can be detected upon click reaction with an azide-coupled reporter. Fluorescent alkyne lipid imaging crucially depends on appropriate azide reporters and labeling protocols that allow for an efficient click reaction and therefore a sensitive detection. We synthesized several azide reporters with different spacer components and tested their suitability for alkyne lipid imaging in fixed cells. The implementation of a copper-chelating picolyl moiety into fluorescent or biotin-based azide reagents strongly increased the sensitivity of the imaging routine. We demonstrate the applicability and evaluate the performance of this approach using different lipid classes and experimental setups. As azide picolyl reporters allow for reduced copper catalyst concentrations, they also enable coimaging of alkyne lipids with multiple fluorescent proteins including enhanced green fluorescent protein. Alternatively, and as we also show, microscopy of alkyne lipids can be combined with protein detection by immunocytochemistry. In summary, we present a robust, sensitive, and highly versatile protocol for the labeling of alkyne lipids with azide-coupled reporters for fluorescence microscopy that can be combined with different protein detection and imaging techniques.
journal_name
J Lipid Resjournal_title
Journal of lipid researchauthors
Gaebler A,Penno A,Kuerschner L,Thiele Cdoi
10.1194/jlr.D070565subject
Has Abstractpub_date
2016-10-01 00:00:00pages
1934-1947issue
10eissn
0022-2275issn
1539-7262pii
jlr.D070565journal_volume
57pub_type
杂志文章abstract：:Scavenger receptor class B type I (SR-BI) and ABCA1 are structurally dissimilar cell surface proteins that play key roles in HDL metabolism. SR-BI is a receptor that binds HDL with high affinity and mediates both the selective lipid uptake of cholesteryl esters from lipid-rich HDL to cells and the efflux of unesterifi...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M300358-JLR200
更新日期：2004-07-01 00:00:00
abstract：:Apolipoprotein AV (apoAV) overexpression causes a decrease in plasma triglyceride (TG) levels, while deficiency of apoAV causes hypertriglyceridemia in both men and mice. However, contrary to what would be expected, plasma apoAV and TG levels in humans are positively correlated. To address this apparent paradox, we de...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M800551-JLR200
更新日期：2009-05-01 00:00:00
abstract：:Two new 6-hydroxylated bile acids, 3 beta, 6 alpha, 12 alpha- and 3 beta, 6 beta, 12 alpha-trihydroxy-5 beta-cholanoic acids, were synthesized from deoxycholic acid. In addition, their C-3 epimers, 3 alpha, 6 alpha, 12 alpha- and 3 alpha, 6 beta, 12 alpha-trihydroxy acids, were prepared by a new route. The principal r...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1991-04-01 00:00:00
abstract：:To consider possible interaction of the phospholipid membrane with calcium ions, crystal structures of calcium dl-alpha- and beta-glycerophosphates (alpha- and beta-CaGs, respectively) were investigated by X-ray diffraction methods. After many attempts, relatively large single crystals of beta-CaG were prepared from t...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1992-07-01 00:00:00
abstract：:Characterization of the chemical form and stereo-specificity of the fatty acid derivatives of arachidonic and linoleic acid in psoriatic epidermis is needed to define the enzymatic origin of these compounds and their possible role in pathogenesis. In an analysis of psoriatic skin scales, both free and esterified 13-hy...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1990-01-01 00:00:00
abstract：:Semi-purified diets supplemented with either a high linoleate (n-6) (safflower) oil or a high alpha-linolenate (n-3) (perilla) oil were fed to mouse mothers and their offspring through 6 weeks of age. The proportions of n-3 and n-6 highly unsaturated fatty acids in brain phospholipids reflected the n-3/n-6 balance of ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1993-02-01 00:00:00
abstract：:Oxylipins are potent lipid mediators involved in a variety of physiological processes. Their profiling has the potential to provide a wealth of information regarding human health and disease and is a promising technology for translation into clinical applications. However, results generated by independent groups are r...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.RA120000991
更新日期：2020-11-01 00:00:00
abstract：:Sonication of rat fat cells caused an increase in lipolysis in the absence of lipolytic stimulants (basal lipolysis) and loss of epinephrine responsiveness. Sonication of endogenous lipid droplets from fat cells also induced an increase in lipolysis in the presence of hormone-sensitive lipase (HSL) and loss of epineph...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1994-01-01 00:00:00
abstract：:As 5-lipoxygenase (5-LO) is an emerging target in obesity and insulin resistance, we have investigated whether this arachidonate pathway is also implicated in the progression of obesity-related fatty liver disease. Our results show that 5-LO activity and 5-LO-derived product levels are significantly elevated in the li...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M800101-JLR200
更新日期：2008-12-01 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：:Immunological tolerance is one of the fundamental aspects of the immune system. The CD4(+)CD25(+) regulatory T (Treg) cells have emerged as key players in the development of tolerance to self and foreign antigens. However, little is known about the endogenous factors and mechanisms controlling their suppressive capaci...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M900101-JLR200
更新日期：2009-12-01 00:00:00
abstract：:apoE is a multi-functional protein expressed in several cell types and in several organs. It is highly expressed in adipose tissue, where it is important for modulating adipocyte lipid flux and gene expression in isolated adipocytes. In order to investigate a potential systemic role for apoE that is produced in adipos...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M050567
更新日期：2015-02-01 00:00:00
abstract：:Five samples from three different types of 1500-year-old Mediterranean amphorae, as well as from a contemporary oil lamp found in the same deposit, were analyzed for the presence of lipid residues. Each sample of finely ground amphorae powder weighed 1-2 g. The abundance of interfering secondary products makes thin-la...
journal_title：Journal of lipid research
pub_type： 历史文章,杂志文章
doi：
更新日期：1981-07-01 00:00:00
abstract：:C26 Bile of the 24-nor-5beta-cholestane series were prepared starting from methyl cholate. A Grignard reaction of methyl magnesium iodide with methyl cholate yielded 24-nor-5beta-cholestane-3alpha, 7alpha, 12alpha, 25-tetrol which was dehydrated TO FORM a mixture of 24-nor-5beta-cholest-23-ene-3alpha, 7alpha, 12 alpha...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1976-09-01 00:00:00
abstract：:Acetyl-L-carnitine is known as a reservoir of activated acetyl units and as a modulator of metabolic function. The objective of this study was to quantify the fate of the acetyl moiety of acetyl-L-carnitine in lipogenic pathways. Lipogenesis was studied in an adipocyte model, differentiated 3T3-L1 cells, and a hepatom...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1997-07-01 00:00:00
abstract：:Adiponectin, an adipocyte-derived protein, has anti-atherogenic and anti-diabetic effects, but how it confers the anti-atherogenic effects is not well understood. To study the anti-atherogenic mechanisms of adiponectin, we examined whether it interacts with atherogenic low-density lipoprotein (LDL) to attenuate LDL's ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.RA120000767
更新日期：2020-11-03 00:00:00
abstract：:The cellular content of total and individual phospholipids and gangliosides was measured in fibroblasts cultured from four normal subjects, three patients with lysosomal lipid storage diseases, and two subjects with homozygous familial hypercholesterolemia. Measurements were made on cells grown in medium containing fe...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1978-03-01 00:00:00
abstract：:Adipose tissue was shown to contain 0.6-1.6 mg of cholesterol per gram wet weight. When expressed per unit of protein or organ mass, fat tissue contains more cholesterol than most other organs or membranes. The cholesterol content of fat tissue increased with the age and weight of the rat. Over 95% of adipose tissue s...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1973-05-01 00:00:00
abstract：:Two assays for the quantitative measurement of HMG-CoA reductase, the major regulatory enzyme in hepatic cholesterol biosynthesis, are described. The first of these procedures employs thin-layer chromatography (TLC) of the reaction product on plastic-backed silica gel G strips impregnated with ammonium carbonate. The ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1977-05-01 00:00:00
abstract：:Acute myeloid leukemia (AML) is the most common acute leukemia in adults. More than half of older AML patients fail to respond to cytotoxic chemotherapy, and most responders relapse with drug-resistant disease. Failure to achieve complete remission can be partly attributed to the drug resistance advantage of AML blast...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M091876
更新日期：2019-06-01 00:00:00
abstract：:Chronic inflammation contributes to cardiovascular disease. Increased levels of the inflammatory cytokine, TNF-α, are often present in conditions associated with cardiovascular disease risk, and TNF-α induces a number of pro-atherogenic effects in macrovascular endothelial cells, including expression of adhesion molec...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.RA119000256
更新日期：2020-01-01 00:00:00
abstract：:The human apolipoprotein C-II locus exhibits genetically determined structural polymorphism in United States and African blacks. In the present study, we have investigated the effect of the apoC-II polymorphism on quantitative serum levels of total cholesterol, total high density lipoprotein (HDL) cholesterol, cholest...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1989-09-01 00:00:00
abstract：:We have studied the recognition and uptake of acid lipase by human fibroblasts in order to determine requirements for localization and function of the enzyme in lysosomes. Our approach was based on evidence that a number of acid hydrolases involved in mucopolysaccharide metabolism are secreted from cultured fibroblast...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：
更新日期：1982-01-01 00:00:00
abstract：:Bile acids are physiological detergents that generate bile flow and facilitate intestinal absorption and transport of lipids, nutrients, and vitamins. Bile acids also are signaling molecules and inflammatory agents that rapidly activate nuclear receptors and cell signaling pathways that regulate lipid, glucose, and en...
journal_title：Journal of lipid research
pub_type： 杂志文章,评审
doi：10.1194/jlr.R900010-JLR200
更新日期：2009-10-01 00:00:00
abstract：:Black tea is one of the world's most popular beverages, and its health-promoting effects have been intensively investigated. The antiobesity and hypolipidemic effects of black tea have attracted increasing interest, but the mechanisms underlying these phenomena remain unclear. In the present study, the black tea major...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M700128-JLR200
更新日期：2007-11-01 00:00:00
abstract：:In breast cancer, 17β-estradiol (E2) plays critical roles mainly by binding to its canonical receptor, estrogen receptor (ER) α66, and eliciting genomic effects. E2 also triggers rapid, nongenomic responses. E2 activates sphingosine kinase 1 (SphK1), increasing sphingosine-1-phosphate (S1P) that binds to its receptors...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M085191
更新日期：2018-12-01 00:00:00
abstract：:Apolipoprotein B (apoB) is a nonexchangeable apolipoprotein. During lipoprotein assembly, it recruits phospholipids and triacylglycerols (TAG) into TAG-rich lipoprotein particles. It remains bound to secreted lipoproteins during lipid metabolism in plasma. The beta1 region (residues 827-1880) of apoB has a high amphip...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M900040-JLR200
更新日期：2009-07-01 00:00:00
abstract：:A deficiency of lysosomal phospholipase A2 (LPLA2) causes macrophage-associated phospholipidosis, suggesting that the enzyme is important in the lipid catabolism. Because LPLA2 is secreted by macrophages, extracellular LPLA2 activity may potentially reflect a change in macrophage activation. In this report, the detect...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.D007146
更新日期：2010-08-01 00:00:00
abstract：:Gangliosides and sulfatides (STs) are acidic glycosphingolipids (GSLs) that have one or more sialic acids or sulfate substituents, in addition to neutral sugars, attached to the C-1 hydroxyl group of the ceramide long chain base. TLC is a widely employed and convenient technique for separation and characterization of ...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.D046128
更新日期：2014-04-01 00:00:00
abstract：:Apolipoprotein (apo)E is well established as a secreted protein that plays an important role in systemic lipoprotein metabolism and vascular wall homeostasis. Recently, endogenous expression of apoE in adipocytes has been shown to play an important role in adipocyte lipoprotein metabolism and gene expression consisten...
journal_title：Journal of lipid research
pub_type： 杂志文章
doi：10.1194/jlr.M011809
更新日期：2011-03-01 00:00:00
