Quantitative analysis of phospholipids by thin‐layer chromatography and phosphorus analysis of spots

Rouser, George; Siakotos, A. N.; Fleischer, Sidney

doi:10.1007/bf02668129

Cited by 1,557 publications

(555 citation statements)

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“…The lower phase from the lipid extraction was dried under a stream of nitrogen gas, and lipids were separated by thin-layer chromatography as described above. The bands corresponding to phosphatidylserine, phosphatidylethanolamine and phosphatidylcholine were scraped from the plate, and the amount of phospholipid was determined by measuring the amount of inorganic phosphorus 47 . It is noteworthy that, in this solvent system, the phosphatidylcholine band slightly overlaps that corresponding to phosphatidylinositol.…”

Section: Metabolic Labeling Of Phosphatidylserine and Phosphatidylethmentioning

confidence: 99%

Gain-of-function mutations in the phosphatidylserine synthase 1 (PTDSS1) gene cause Lenz-Majewski syndrome

Jenkins²,

et al. 2013

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Lenz-Majewski syndrome (LMS) is a syndrome of intellectual disability and multiple congenital anomalies that features generalized craniotubular hyperostosis. By using whole-exome sequencing and selecting variants consistent with the predicted dominant de novo etiology of LMS, we identified causative heterozygous missense mutations in PTDSS1, which encodes phosphatidylserine synthase (PSS ). PSS is one of two enzymes involved in the production of phosphatidylserine. Phosphatidylserine synthesis was increased in intact fibroblasts from affected individuals, and end-product inhibition of PSS by phosphatidylserine was markedly reduced. Therefore, these mutations cause a gain-of-function effect associated with regulatory dysfunction of PSS . We have identified LMS as the first human disease, to our knowledge, caused by disrupted phosphatidylserine metabolism. Our results point to an unexplored link between phosphatidylserine synthesis and bone metabolism.

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Section: Metabolic Labeling Of Phosphatidylserine and Phosphatidylethmentioning

confidence: 99%

Gain-of-function mutations in the phosphatidylserine synthase 1 (PTDSS1) gene cause Lenz-Majewski syndrome

Jenkins²,

et al. 2013

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“…The wild-type membrane fraction showed significant LPIAT activity by using sn-1-lysoPI as a substrate (black plus gray bars in the Supplemental Figure S1A), and the enzyme activity was reduced to ϳ65% in the membranes of mboa-7 mutants. Because sn-1-lysoPI is known to easily isomerize to form sn-2-lysoPI (see Supplemental Figure S1B; Rouser et al, 1966), it was possible that 14 C-acyl donor was incorporated into sn-2-lysoPI and sn-1-lysoPI. To check the positional specificity, [ 14 C]PI produced after the incubation was treated again with bee venom phospholipase A 2 (for details, see the legend to Supplemental Figure S1).…”

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confidence: 99%

Caenorhabditis elegans mboa-7, a Member of the MBOAT Family, Is Required for Selective Incorporation of Polyunsaturated Fatty Acids into Phosphatidylinositol

Lee

Inoué

Imae

et al. 2008

MBoC

125

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Phosphatidylinositol (PI) is a component of membrane phospholipids, and it functions both as a signaling molecule and as a compartment-specific localization signal in the form of polyphosphoinositides. Arachidonic acid (AA) is the predominant fatty acid in the sn-2 position of PI in mammals. LysoPI acyltransferase (LPIAT) is thought to catalyze formation of AA-containing PI; however, the gene that encodes this enzyme has not yet been identified. In this study, we established a screening system to identify genes required for use of exogenous polyunsaturated fatty acids (PUFAs) in Caenorhabditis elegans. In C. elegans, eicosapentaenoic acid (EPA) instead of AA is the predominant fatty acid in PI. We showed that an uncharacterized gene, which we named mboa-7, is required for incorporation of PUFAs into PI. Incorporation of exogenous PUFA into PI of the living worms and LPIAT activity in the microsomes were greatly reduced in mboa-7 mutants. Furthermore, the membrane fractions of transgenic worms expressing recombinant MBOA-7 and its human homologue exhibited remarkably increased LPIAT activity. mboa-7 encodes a member of the membrane-bound O-acyltransferase family, suggesting that mboa-7 is LPIAT. Finally, mboa-7 mutants had significantly lower EPA levels in PI, and they exhibited larval arrest and egg-laying defects. INTRODUCTIONVarious kinds of fatty acids are distributed in membrane phospholipids in mammalian cells and tissues (Lands and Crawford, 1976;Holub and Kuksis, 1978;MacDonald and Sprecher, 1991). The fatty acyl residues of individual phospholipids seem to be under strict metabolic regulation. In general, saturated fatty acids are esterified at the sn-1 position, whereas polyunsaturated fatty acids (PUFAs), such as arachidonic acid (AA), are commonly found at the sn-2 position. Three-fourths or more of the phosphatidylinositol (PI) fraction in rat liver and brain constitutes the 1-stearoyl-2-arachidonoyl species (Holub and Kuksis, 1971;Baker and Thompson, 1972). In contrast, the total pool of precursor phosphatidic acid (PA) in rat liver and brain has a fatty acid composition that does not resemble that of PI, showing a low AA content (Possmayer et al., 1969;Akesson et al., 1970;Baker and Thompson, 1972). Selectivity could be expressed during de novo synthesis at the level of formation of cytidine 5Ј-diphosphate (CDP)-diacylglycerol from PA and cytidine 5Ј-triphosphate or in the use of CDP-diacylglycerol in the final reaction. In fact, CDP-diacylglycerol synthase, which prefers 1-stearoyl-2-arachidonoyl PA as a substrate in vitro, has been cloned, although expression is restricted to testis, retina, and brain (Saito et al., 1997).An alternative mechanism for species selection has been proposed on the basis of turnover studies in rat brain in vivo (Baker and Thompson, 1972). [ 3 H]AA and [ 14 C]glycerol injected intracerebrally were incorporated almost exclusively into brain phospholipids. Comparison of PI and PA radioactivity suggest that the initial flux of AA into PI was independent of de novo synthesi...

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“…LPC, SM, PC, PI1PS (separation of PI from PS is technically challenging) and PE were stripped from each area. The contents of LPC, SM, PC, PI1PS, PE, and total PL in the samples were measured with phosphorus quantification (21). The contents of each PL molecular class were calculated according to the phosphorus contents using the molecular weight of each PL class.…”

Section: Meal Samplesmentioning

confidence: 99%

Quantities of Phospholipid Molecular Classes in Japanese Meals and Prediction of Their Sources by Multiple Regression Analysis

Shirouchi

Yamanaka

Tanaka

et al. 2018

Journal of Nutritional Science and Vitaminology

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Quantitative analysis of phospholipids by thin‐layer chromatography and phosphorus analysis of spots

Cited by 1,557 publications

References 2 publications

Gain-of-function mutations in the phosphatidylserine synthase 1 (PTDSS1) gene cause Lenz-Majewski syndrome

Gain-of-function mutations in the phosphatidylserine synthase 1 (PTDSS1) gene cause Lenz-Majewski syndrome

Caenorhabditis elegans mboa-7, a Member of the MBOAT Family, Is Required for Selective Incorporation of Polyunsaturated Fatty Acids into Phosphatidylinositol

Quantities of Phospholipid Molecular Classes in Japanese Meals and Prediction of Their Sources by Multiple Regression Analysis

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