A two-dimensional thin-layer chromatographic system for phospholipid separation the analysis of yeast phospholipids

Getz, Godfrey S.; Jakovcic, Smilja; Heywood, Joanna; Frank, J.; Rabinowitz, Murray

doi:10.1016/0005-2760(70)90007-x

Cited by 97 publications

(42 citation statements)

References 26 publications

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“…Comparison with data from other studies is made difficult not only by there being differences in culture age and phase, but also by the fact that different media are used which can affect lipid content (and composition) markedly (e.g. see Getz et al, 1970). Phosphatidylcholine, phosphatidylethanolaine and phosphatidylinositol were the most abundant phospholipids in our strains, as observed previously in other C .…”

Section: Discussionsupporting

confidence: 53%

The Lipid Composition of Azole-sensitive and Azole-resistant Strains of Candida albicans

Hitchcock¹,

Barrett‐Bee²,

Russell³

1986

Microbiology

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The lipid compositions of two azole-sensitive (A and B2630) and two azole-resistant (AD and KB) strains of the opportunistic fungal pathogen Candida albicans were studied by using several lipid extraction procedures: no differences were observed between the lipid content or total phospholipid/neutral lipid ratios of the four strains. All contained phosphatidylethanolamine, phosp hatidylc holine, phosphatid ylinosi to1 and p hosp hat id ylserine as major phospholipids, with smaller amounts of phosphatidylglycerol and diphosphatidylglycerol ; the relative proportions of these lipids differed between all four strains. The fatty acid composition of each major phospholipid within each strain differed, and there were also interstrain differences. A marked effect of culture growth phase in batch culture on lipid composition was observed. The major neutral lipids in each strain were triacylglycerol, non-esterified sterol and non-esterified fatty acid. The fatty acid compositions of the three fatty-acid-containing neutral lipids were distinct from each other and the phospholipids, and there were also interstrain differences. All strains possessed (1yso)phospholipase activity, which was nonspecific. The proportions of triacylglycerol and non-esterified fatty acid did not vary between strains, but the azole-resistant strains AD and KB contained more non-esterified sterol, giving them a phospholipid/sterol ratio approximately half that of azole-sensitive strains. There appeared to be a relationship between the phospholipid/sterol ratio of exponentially growing sensitive strains and their ability to take up azole; this did not extend to the resistant strains, which either did not take up azole (AD and KB) or took it up at a faster rate (Darlington) than sensitive strains. INTRODUCTIONCandida albicans is a widespread and troublesome opportunistic pathogen that causes a variety of superficial and deep-seated mycoses (Odds, 1979). Of the relatively few antifungal antibiotics available, those most commonly used to treat candidosis are the polyenes and the more recently introduced imidazoles (Speller, 1980). Polyenes work by complexing with membrane sterols (Hamilton-Miller, 1973). The primary action of imidazoles is probably inhibition of ergosterol synthesis that leads to an accumulation of 14a-methyl sterols which disrupt membrane structure and function (Van den Bossche et ul., 1982, 1983). However, imidazole actions are complex: they also inhibit a number of other yeast (and mammalian) membrane-bound enzymes (Uno et al., Mason et al., 1985), and at higher concentrations some of them affect yeast membranes by direct interaction with lipids (Cope, 1980; Brasseur et al., 1983).Clinical isolates of C. albicans vary considerably in their sensitivities to imidazoles (e.g. see Ryley et al., 1984), and azole-resistant strains have been isolated (Holt & Azmi, 1978 ;Horsburgh & Kirkpatrick, 1983; Warnock et al., 1983). The failure of two of these resistant strains (AD and , 1984). This might be due to an altered membrane lipid compos...

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Section: Discussionsupporting

confidence: 53%

The Lipid Composition of Azole-sensitive and Azole-resistant Strains of Candida albicans

Hitchcock¹,

Barrett‐Bee²,

Russell³

1986

Microbiology

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“…Analysis of 10 consecutive preparations of islets, after labeling with 32P and washing, indicated that 68.8 ± 1.6% 1 of the radioactivity present was acid-soluble, 7.2 +0.7% had been incorporated into the phospholipid fraction and the remainder was in the nucleic acid and phosphoprotein residue. Two-dimensional thin-layer chromatography (14) showed that a majority (73.2±1.7%o) of the phospholipid labeling was in the phosphatidylinositol fraction.…”

Section: Methodsmentioning

confidence: 99%

Rapid Transient Efflux of Phosphate Ions from Pancreatic Islets as an Early Action of Insulin Secretagogues

Freinkel¹,

Younsi²,

J³

et al. 1974

J. Clin. Invest.

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A B S T R A C T Anionic fluxes during the membrane realignments of stimulated insulin release have not been characterized previously although cations have been implicated in stimulus-secretion coupling. We have shown that a limited packet pulse of phosphate release ("phosphate flush") begins at the same time that the first phase of insulin secretion may occur. To demonstrate this phenomenon, we have prelabeled islets, obtained from rat pancreas by collagenase digestions, by incubation with [8'P]orthophosphate. When such prelabeled islets are perifused with Krebs-Ringer bicarbonate containing 0.5 mg/ml D-glucose, a basal rate of efflux of radioactivity is established; transfer to perifusates containing 3.0 mg/ml n-glucose elicits an increased 'P efflux within 1-2 min to peak values which are 7-to 21-fold greater than basal. The total duration of this "phosphate flush" approximates 10 min and exceeds the duration of the first phase of stimulated insulin secretion. With lesser concentrations of glucose, the flush exhibits dose-response relationships, and with 3 mg/ml glucose, a second flush can be elicited by restoring basal conditions and stimulating anew with 3 mg/ml glucose. The phenomenon is highly specific and can be reduplicated by other secretagogues (L-leucine) or sugars (D-mannose) which are also known to elicit insulin release but not by sugars which fail to affect insulin secretion (D-galactose, D-fructose, i-inositol, L-glucose). The efflux of radioactivity consists entirely of ["P]orthophosThis work was presented in part at

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“…Cultures were grown aerobically on enriched media as described by Getz et al (1970) . In some experiments 2%Jo galactose or 2 0/o Na lactate replaced glucose as the carbon source in the medium.…”

Section: Culture Mediamentioning

confidence: 99%

Cardiolipin Content of Wild Type and Mutant Yeasts in Relation to Mitochondrial Function and Development

Jakovcic

Getz

Rabinowitz

et al. 1971

The Journal of Cell Biology

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The phospholipid composition of various strains of the yeast, Saccharomyces cerevisiae, and several of their derived mitochondrial mutants grown under conditions designed to induce variations in the complement of mitochondrial membranes has been examined . Wild type and petite (cytoplasmic respiratory deficient) yeasts were fractionated into various subcellular fractions, which were monitored by electron microscopy and analyzed for cytochrome oxidase (in wild type) and phospholipid composition . 90% or more of the phospholipid, cardiolipin was found in the mitochondrial membranes of wild type and petite yeast . Cardiolipin content differed markedly under various growth conditions . Stationary yeast grown in glucose had better developed mitochondria and more cardiolipin than repressed log phase yeast . Aerobic yeast contained more cardiolipin than anaerobic yeast . Respiration-deficient cytoplasmic mitochondrial mutants, both suppressive and neutral, contained less cardiolipin than corresponding wild types . A chromosomal mutant lacking respiratory function had normal cardiolipin content . Log phase cells grown in galactose and lactate, which do not readily repress the development of mitochondrial membranes, contained as much cardiolipin as stationary phase cells grown in glucose . Cytoplasmic mitochondrial mutants respond to changes in the glucose concentration of the growth medium by variations in their cardiolipin content in the same way as wild type yeast does under similar growth conditions. It is concluded that cardiolipin content of yeast is correlated with, and is a good indicator of, the state of development of mitochondrial membrane .

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A two-dimensional thin-layer chromatographic system for phospholipid separation the analysis of yeast phospholipids

Cited by 97 publications

References 26 publications

The Lipid Composition of Azole-sensitive and Azole-resistant Strains of Candida albicans

The Lipid Composition of Azole-sensitive and Azole-resistant Strains of Candida albicans

Rapid Transient Efflux of Phosphate Ions from Pancreatic Islets as an Early Action of Insulin Secretagogues

Cardiolipin Content of Wild Type and Mutant Yeasts in Relation to Mitochondrial Function and Development

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