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Methods in Molecular Biology (Clifton,... 2019HPLC-MS/MS has enabled the quantitative analysis of complex mixtures of lipid molecular species. Several separate analyses, using methods that have been optimized for...
HPLC-MS/MS has enabled the quantitative analysis of complex mixtures of lipid molecular species. Several separate analyses, using methods that have been optimized for individual lipid classes, provide good lipidomic profiles, but may not be desirable for laboratories constrained by available instrumentation and wanting a higher throughput. Here we describe two methods using binary gradient HiLiC HPLC and triple quadrupole MS that together provide a lipidomic profile for lipids of interest in type 2 diabetes research. Methods for analysis of molecular species of diacylglycerol, ceramide, dihydroceramide, sphingosine, glucosyl- and lactosylceramide, sphingomyelin, and acylcarnitine from skeletal muscle and primary culture cells are described.
Topics: Chromatography, High Pressure Liquid; Diabetes Mellitus, Type 2; Diglycerides; Humans; Muscle, Skeletal; Sphingolipids; Tandem Mass Spectrometry
PubMed: 31119661
DOI: 10.1007/978-1-4939-9236-2_9 -
International Journal of Biological... Nov 2023Konjac glucomannan (KGM) combined with hydroxypropyl methyl cellulose was used to fabricate diacylglycerol oleogels (DGOs) through the emulsion-templated method, and... (Comparative Study)
Comparative Study
Konjac glucomannan (KGM) combined with hydroxypropyl methyl cellulose was used to fabricate diacylglycerol oleogels (DGOs) through the emulsion-templated method, and compared with triacylglycerol oleogels (TGOs). The appearance and microstructure results showed that stable emulsions and oleogels could be formed in the presence of 0.2-0.6 wt% KGM. Higher KGM concentrations resulted in a stronger gel structure in oleogels, whose thixotropic recovery percentages were 50.45-75.20 %. From LF-NMR determination, the higher concentration of KGM presented earlier transverse relaxation (T) time, and the T parameters of DGOs were higher than that of TGOs. Texture and oil loss analysis indicated that the mechanical strength and oil holding ability of DGOs were slightly lower than those of TGOs. This study demonstrated the advantages of biopolymers as thickening agents for obtaining stable emulsion and oleogels. The specific characteristics of DGOs distinguished from TGOs should be attributed to their different properties (unsaturation, viscosity, polarity, etc.) between the liquid oils.
Topics: Mannans; Emulsions; Triglycerides; Organic Chemicals; Diglycerides; Viscosity
PubMed: 37598830
DOI: 10.1016/j.ijbiomac.2023.126420 -
PeerJ 2023Olive ( L.) oil accumulate more diacylglycerols (DAG) than mostly vegetable oils. Unsaturated fatty acids-enriched DAG consumption enhanced wellness in subjects....
BACKGROUND
Olive ( L.) oil accumulate more diacylglycerols (DAG) than mostly vegetable oils. Unsaturated fatty acids-enriched DAG consumption enhanced wellness in subjects. However, the mechanism of DAG accumulation is not yet fully understood.
METHODS
In this study, gene network of DAG accumulation and fatty acid composition in the two olive mesocarps ("Chenggu 32" (CG) and "Koroneiki" (QJ)) were investigated by integrating lipidome and transcriptome techniques.
RESULTS
A total of 1,408 lipid molecules were identified by lipidomic analysis in olive mesocarp, of which DAG (DAG36:3, DAG36:4 and DAG36:5) showed higher content, and triacylglycerols (TAG54:3, TAG54:4) exhibited opposite trend in CG. Specifically, DAG was rich in polyunsaturated fatty acids (especially C18:2) at the sn-2 position, which was inconsistent with TAG at the same positions (Primarily C18:1). Transcriptomic analysis revealed that phospholipase C (NPC, EC 3.1.4.3) were up-regulated relative to QJ, whereas diacylglycerol kinase (ATP) (DGK, EC 2.7.1.107), diacylglycerol acyltransferase (DGAT, EC 2.3.1.20), and phospholipid: diacylglycerol acyltransferase (PDAT, EC 2.3.1.158) were down-regulated.
CONCLUSION
We speculated that the non-acyl coenzyme A pathway played a significant role in DAG biosynthesis. Additionally, fatty acyl-ACP thioesterase B (FATB, EC 3.1.2.14), stearoyl [acyl-carrier-protein] 9-desaturase (SAD, EC 1.14.19.2) and omega-6 fatty acid desaturase (FAD2, EC 1.14.19.6) were highly expressed in CG and may be involved in regulating fatty acid composition. Meanwhile, phospholipase A1 (LCAT, EC 3.1.1.32) involved in the acyl editing reaction facilitated PUFA linkage at the sn-2 position of DAG. Our findings provide novel insights to increase the DAG content, improve the fatty acid composition of olive oil, and identify candidate genes for the production of DAG-rich oils.
Topics: Humans; Olea; Lipidomics; Diacylglycerol O-Acyltransferase; Diglycerides; Transcriptome; Fatty Acids; Fatty Acids, Unsaturated
PubMed: 37583911
DOI: 10.7717/peerj.15724 -
Journal of Medicinal Chemistry May 2014To explore the feasibility of developing ligands targeted to the atypical C1 domains of protein kinase C ζ and ι, we have prepared diacylglycerol lactones substituted...
To explore the feasibility of developing ligands targeted to the atypical C1 domains of protein kinase C ζ and ι, we have prepared diacylglycerol lactones substituted with hydrophilic groups on their side chains, which potentially could interact with the arginine residues that distinguish the atypical C1 domains of PKCζ and PKCι from typical C1 domains, and we have measured their binding to mutated versions of the C1b domain of PKCδ that incorporate one or more of these arginine residues. The most selective of the diacylglycerol lactones showed only a 10-fold reduction in binding affinity with the triple arginine mutant (N7R/S10R/L20R) compared to the wild-type, whereas phorbol 12,13-dibutyrate showed a 6000-fold loss of affinity. Molecular modeling confirms that these ligands are indeed able to interact with the arginine residues. Our results show that dramatic changes in selectivity can be obtained through appropriate substitution of diacylglycerol lactones.
Topics: Amino Acid Sequence; Diglycerides; Lactones; Magnetic Resonance Spectroscopy; Models, Molecular; Molecular Sequence Data; Protein Kinase C; Sequence Homology, Amino Acid; Spectrometry, Mass, Fast Atom Bombardment
PubMed: 24684293
DOI: 10.1021/jm500165n -
Methods in Molecular Biology (Clifton,... 1998
Review
Topics: Animals; Chromatography, High Pressure Liquid; Diglycerides; Fatty Acids; Phospholipids
PubMed: 10427569
DOI: 10.1385/0-89603-491-7:255 -
Cell Chemical Biology Feb 2018Diacylglycerol-sensitive transient receptor potential (TRP) channels play crucial roles in a wide variety of biological processes and systems, but their activation...
Diacylglycerol-sensitive transient receptor potential (TRP) channels play crucial roles in a wide variety of biological processes and systems, but their activation mechanism is not well understood. We describe an optical toolkit by which activation and deactivation of these ion channels can be controlled with unprecedented speed and precision through light stimuli. We show that the photoswitchable diacylglycerols PhoDAG-1 and PhoDAG-3 enable rapid photoactivation of two DAG-sensitive TRP channels, Trpc2 and TRPC6, upon stimulation with UV-A light, whereas exposure to blue light terminates channel activation. PhoDAG photoconversion can be applied in heterologous expression systems, in native cells, and even in mammalian tissue slices. Combined laser scanning-controlled photoswitching and Ca imaging enables both large-scale mapping of TRP channel-mediated neuronal activation and localized mapping in small cellular compartments. Light-switchable PhoDAGs provide an important advance to explore the pathophysiological relevance of DAG-sensitive TRP channels in the maintenance of body homeostasis.
Topics: Animals; Diglycerides; Humans; Mice; Mice, Inbred C57BL; Molecular Structure; Photochemical Processes; Transient Receptor Potential Channels
PubMed: 29276045
DOI: 10.1016/j.chembiol.2017.11.008 -
Methods in Molecular Biology (Clifton,... 2023The endocannabinoid 2-arachidonoylglycerol (2-AG) exerts its physiological action by binding to and functionally activating type-1 (CB) and type-2 (CB) cannabinoid...
The endocannabinoid 2-arachidonoylglycerol (2-AG) exerts its physiological action by binding to and functionally activating type-1 (CB) and type-2 (CB) cannabinoid receptors. It is thought to be produced through the action of sn-1 selective diacylglycerol lipase (DAGL) that catalyzes 2-AG biosynthesis from sn-2-arachidonate-containing diacylglycerols. Different methodological approaches for measuring DAGL activity in biological samples are now available. Here, a highly sensitive radiometric assay to assess DAGL activity, by using 1-oleoyl[1-C]-2-arachidonoylglycerol as the substrate, is reported. All the steps required to perform lipid extraction, fractionation by thin-layer chromatography (TLC), and quantification of radiolabeled [C]-oleic acid via scintillation counting are described in detail.
Topics: Diglycerides; Endocannabinoids; Lipoprotein Lipase; Oleic Acid; Receptors, Cannabinoid
PubMed: 36152195
DOI: 10.1007/978-1-0716-2728-0_23 -
Journal of Lipid Research Jun 2000alpha-Cyclodextrins are water-soluble cyclic hexamers of glucose units with hydrophobic cavities capable of solubilizing lipophiles. Incubating alpha-cyclodextrin with...
alpha-Cyclodextrins are water-soluble cyclic hexamers of glucose units with hydrophobic cavities capable of solubilizing lipophiles. Incubating alpha-cyclodextrin with high density lipophorin from Manduca sexta or Bombyx mori resulted in a cloudy, turbid solution. Centrifugation separated a pale yellowish precipitate. Thin-layer chromatography analysis of the lipid extract of the precipitate showed that the major lipid was diacylglycerol, while KBr density gradient analysis of the supernatant demonstrated the presence of a lipid-depleted very high density lipophorin. Transfer of diacylglycerol from lipophorin to cyclodextrin was specific to alpha-cyclodextrin and was not observed with beta- or gamma-cyclodextrins. pH had no effect on diacylglycerol transfer to alpha-cyclodextrin. However, the transfer was strongly dependent on the concentration of alpha-cyclodextrin and temperature. Increasing the concentration of alpha-cyclodextrin in the incubation mixture was associated with the formation of increasingly higher density lipophorins. Thus, at 20, 30, and 40 mm alpha-cyclodextrin, the density of B. mori lipophorin increased from 1.107 g/ml to 1.123, 1. 148, and 1.181 g/ml, respectively. At concentrations greater than 40 mm, alpha-cyclodextrin had no further effect on the density of lipophorin. alpha-Cyclodextrin removed at most 83;-87% of the diacylglycerol present in lipophorin. Temperature played an important role in altering the amount of diacylglycerols transferred to alpha-cyclodextrin. At 30 mm alpha-cyclodextrin, the amount of diacylglycerol transferred at different temperatures was 50% at 4 degrees C, 41% at 15 degrees C, 20% at 28 degrees C, and less than 3% at 37 degrees C. We propose that diacylglycerol transfers to alpha-cyclodextrin via an aqueous diffusion pathway and that the driving force for the transfer is the formation of an insoluble alpha-cyclodextrin-diacylglycerol complex.
Topics: Animals; Carrier Proteins; Cyclodextrins; Diglycerides; Hydrogen-Ion Concentration; Insecta; Lipoproteins; Lipoproteins, HDL; Temperature; alpha-Cyclodextrins
PubMed: 10828085
DOI: No ID Found -
Diabetes Research and Clinical Practice Jan 2020Hepatic insulin resistance, induced by fat, occurs before peripheral resistance and leads to prediabetes and diabetes. If insulin resistance is detected earlier,...
AIMS
Hepatic insulin resistance, induced by fat, occurs before peripheral resistance and leads to prediabetes and diabetes. If insulin resistance is detected earlier, lifestyle changes could prevent or delay disease development. Therefore, we analysed lipids in the liver and serum of prediabetic and diabetic mice by MeV TOF-SIMS with a focus on diacylglycerols (DAGs) as the best predictor of (liver) resistance.
METHODS
Glucose impairment was spontaneously developed or induced by HFD in NOD/LtJ mice, and prediabetic and diabetic mice were selected according to their glucose levels. MeV TOF-SIMS was applied to image the lipid distribution in the liver and to relatively quantify lipids related to insulin resistance in both the liver and serum.
RESULTS
The same lipids were detected in the liver and serum but with different intensities between mice. The intensity of DAGs and fatty acids was higher in the diabetic than that in the prediabetic liver. Imaging of liver tissue showed a more compact density of prediabetic (non-fatty) than diabetic liver with DAG remodelling in diabetes. DAGs, which are greatly increased in diabetic serum, were successfully detected and quantified already in prediabetes.
CONCLUSION
MeV TOF-SIMS applied to the serum presents an excellent tool for in vivo monitoring of disease development over time.
Topics: Animals; Blood Chemical Analysis; Diabetes Mellitus, Experimental; Diglycerides; Fatty Acids; Female; Insulin Resistance; Lipid Metabolism; Lipids; Liver; Male; Mice; Mice, Inbred NOD; Prediabetic State; Spectrometry, Mass, Secondary Ion
PubMed: 31866525
DOI: 10.1016/j.diabres.2019.107986 -
European Journal of Histochemistry : EJH 2004The story of nuclear diacyglycerol is proving to be a complex one. Sub-pools of nuclear diglyceride that differ in their metabolism, nuclear localization and temporal... (Review)
Review
The story of nuclear diacyglycerol is proving to be a complex one. Sub-pools of nuclear diglyceride that differ in their metabolism, nuclear localization and temporal regulation have been identified, suggesting potentially diverse signaling functions. One of the great remaining challenges is to assign functional roles to these diverse populations. In the last twenty years great strides have been made toward understanding the character and composition of nuclear DAG. Determining the functions of this nuclear lipid should make the next twenty years interesting indeed.
Topics: Animals; Cell Nucleus; Diacylglycerol Kinase; Diglycerides; Humans; Lipids; Signal Transduction
PubMed: 15145778
DOI: No ID Found