Analysis of membrane lipids by 500 MHz 1H NMR

Sparling, Mary Lee; Zidovetzki, Raphael; Müller, Luciano; Chan, Sunney I.

doi:10.1016/0003-2697(89)90358-8

Cited by 66 publications

(54 citation statements)

References 15 publications

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“…The residue was dissolved in CD 3 OD/CDCl 3 mixture (3 : 2 v/v) and subjected to NMR analysis. 11,12 NMR measurements NMR spectra of lettuce extracts were recorded at 300 K on a Bruker AVANCE AQS600 spectrometer operating at the 1 H frequency of 600.13 MHz and equipped with a Bruker multinuclear z-gradient inverse probehead capable of producing gradients in the z-direction with a strength of 55 G cm 1 . Proton spectra were referenced to methyl group signals at υ D 0.00 ppm of TSPA in D 2 O and to the CH 3 resonance of tetramethylsilane (TMS) in CD 3 OD/CDCl 3 , whereas 13 C spectra were referenced to the anomeric 13 1 H spectra of aqueous extracts were acquired by coadding 512 transients with a recycle delay of 3 s. The residual HDO signal was suppressed using a presaturation with a long single soft pulse at HDO resonance frequency applied during the relaxation delay followed by a strong observation pulse.…”

Section: Introductionmentioning

confidence: 99%

Metabolic profile of lettuce leaves by high-field NMR spectra

Sobolev¹,

Brosio²,

Gianferri³

et al. 2005

Magn. Reson. Chem.

153

138

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A detailed analysis of the proton high-field NMR spectra of aqueous and organic extracts of lettuce leaves is reported for the first time. A combination of COSY, TOCSY, 1 H-13 C HSQC, 1 H-13 C HMBC bidimensional sequences and DOSY was used to assign each spin system and to separate the components of the complex patterns. A large number of water-soluble metabolites belonging to different classes such as carbohydrates, polyols, organic acids and amino acids were fully assigned. Moreover, the complex spectra of metabolites extracted in organic solvents belonging to sterols, fatty acids, diacylglycerophospholipids, galactosyldiacylglycerols, sulpholipids, pheophytins, carotenoids and hydrocarbons were also assigned.

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Section: Introductionmentioning

confidence: 99%

Metabolic profile of lettuce leaves by high-field NMR spectra

Sobolev¹,

Brosio²,

Gianferri³

et al. 2005

Magn. Reson. Chem.

153

138

View full text Add to dashboard Cite

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“…For each lipid class, the spectral assignments were compared to standard spectra of CL, PI, and PG (1,7,48). Diagnostic resonances may be considered to be the CL resonances of the central glycerol head group, sandwiched between two phosphate groups (OPCH 2 CHOHCH 2 OP) ( Table 3 and Fig.…”

mentioning

confidence: 99%

Coordinated Regulation of Cold-Induced Changes in Fatty Acids with Cardiolipin and Phosphatidylglycerol Composition among Phospholipid Species for the Food Pathogen Listeria monocytogenes

Mastronicolis

Arvanitis

Karaliota

et al. 2008

Appl Environ Microbiol

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We present here the structural identification of four phospholipid (Phl) classes in Listeria monocytogenes, the fatty acid (FA) composition for each individual Phl species, and a description of cold-induced FA changes. Cardiolipin (48.5%) and phosphatidylglycerol (18.1%) are dominated by anteiso-FA, and the previously recognized branched FA chain shortening by cold was observed singularly in these Phls. Phosploaminolipid (19.9%) and phosphatidylinositol, (9.1%) are significantly different, containing significant amounts of straightchain FA. These findings are supported by nuclear magnetic resonance analysis.The bacterium Listeria monocytogenes, one of the leading causes of food-borne illness (15), can adapt to cold and salinity (9,10,31,46,53) and can survive even on dust or flakes of organic material (34). Listeria is characterized by Ͼ85% branched-chain fatty acids (FA), anteiso-15:0 (a-15:0) and anteiso-17:0 (a-17:0). Cells grown in the cold (5°C) contain significantly less a-17:0 than those grown at higher temperatures (4,8,37,43,44,57). A cryotolerance element of Listeria is also its ability to use cryoprotectant osmolytes (31), a property shared by other organisms (2, 3, 12, 21, 23). The critical role of odd numbered anteiso-FA in influencing the lower-temperature growth limits has recently been reported (14,19,25). We have reported previously (39) that Listeria spp. can be cold adapted by an increased content (30%) of neutral lipids (NL) among total lipids (TL) and an increase in the a-15:0/a-17:0 FA ratio (FAr) of TL, polar lipids (PL), and NL (10-, 10-, and 7-fold, respectively), as well as by an increase in FAr (38) for each NL subclasses 1,2-diglyceride, 1,3-diglyceride and free FA (6-, 5-, and 8-fold, respectively).This study is a continuation (35, 36) of an investigation for the structural characterization of the major phospholipid (Phl) classes of L. monocytogenes, in order to (i) elucidate the isolation and quantification of each of class (cells at 30°C), (ii) evaluate the cold temperature (5°C) effect on each individual Phl class FA composition, and (iii) confirm the findings by 1 H-nuclear magnetic resonance (NMR) analysis.

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“…Although most of these studies have concentrated on aqueous metabolites, qualitative and quantitative analysis of tissue lipid extracts and biofluids probably would also provide specific clues of diagnostic importance (14)(15)(17)(18)(19)(20). Most of the popular methods involve perchloric acid (PCA) extraction for aqueous soluble metabolites (16,(21)(22)(23)(24) and methanol/chloroform (M/C) extraction for lipid components (25)(26)(27)(28)(29)(30)(31)(32)(33). However, biopsy specimens of human tissues are so small that lipids and aqueous metabolites often have to be studied in the same tissue sample.…”

Section: Introductionmentioning

confidence: 99%

Novel corrective equations for complete estimation of human tissue lipids after their partial destruction by perchloric acid pre‐treatment: high‐resolution ¹H‐NMR‐based study

et al. 2008

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Owing to the small quantity of tissue available in human biopsy specimens, aqueous and lipid components often have to be determined in the same tissue sample. Perchloric acid (PCA) used for the extraction of aqueous metabolites has a deleterious effect on lipid components; the severity of the damage is not known. In this study, human muscle tissue was first treated with PCA to extract aqueous metabolites, and the residue was then used for lipid extraction by conventional methods, i.e. the methods of Folch and Bligh & Dyer and a standardised one using methanol/chloroform (1:3, v/v) used in our laboratory. A (1)H-NMR spectrum was obtained for each lipid extract. Lipid was quantified by measuring the integral area of N(+)-(CH(3))(3) signals of phospholipids (PLs). Triacylglycerol (TG) and cholesterol (CHOL) were quantified using the -CH(2)- signals of glycerol and the C18 methyl signal, respectively. This study shows that prior use of PCA caused marked attenuation of TG, PL, and CHOL. This was confirmed by recovery experiments and observation of the direct effect of PCA on the standard lipid components. On the basis of the quantity of lipid lost in each case, three novel equations (with respect to TG, PL, and CHOL) were derived. Application of these equations to lipid quantities estimated in different pathological tissues after PCA pre-treatment produced values equivalent to those estimated without PCA use. This study conclusively shows that PCA pre-treatment damages all three lipid moieties, TG, PL, and CHOL. When PCA is used in a fixed ratio to the tissue, the lipid damage is also proportional and correctable by statistically derived equations. These equations will be useful in human biopsy specimens where aqueous and lipid components have to be studied using the same tissue sample because of the small quantity available.

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Analysis of membrane lipids by 500 MHz 1H NMR

Cited by 66 publications

References 15 publications

Metabolic profile of lettuce leaves by high-field NMR spectra

Metabolic profile of lettuce leaves by high-field NMR spectra

Coordinated Regulation of Cold-Induced Changes in Fatty Acids with Cardiolipin and Phosphatidylglycerol Composition among Phospholipid Species for the Food Pathogen Listeria monocytogenes

Novel corrective equations for complete estimation of human tissue lipids after their partial destruction by perchloric acid pre‐treatment: high‐resolution ¹H‐NMR‐based study

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Analysis of membrane lipids by 500 MHz 1H NMR

Cited by 66 publications

References 15 publications

Metabolic profile of lettuce leaves by high-field NMR spectra

Metabolic profile of lettuce leaves by high-field NMR spectra

Coordinated Regulation of Cold-Induced Changes in Fatty Acids with Cardiolipin and Phosphatidylglycerol Composition among Phospholipid Species for the Food Pathogen Listeria monocytogenes

Novel corrective equations for complete estimation of human tissue lipids after their partial destruction by perchloric acid pre‐treatment: high‐resolution 1H‐NMR‐based study

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Novel corrective equations for complete estimation of human tissue lipids after their partial destruction by perchloric acid pre‐treatment: high‐resolution ¹H‐NMR‐based study