Turnover of phosphomonoester groups and compartmentation of polyphosphoinositides in human erythrocytes

Müller, Elke; Hegewald, H; Jaroszewicz, K; Cumme, Gerhard A.; Hoppe, H. H.; Frunder, Horst

doi:10.1042/bj2350775

Cited by 53 publications

(37 citation statements)

References 26 publications

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“…This fact could be connected to the maintenance of erythrocyte physiological shape. However, Schwartz et al (1985) suggest that diacylglycerol as an intermediate phosphoinositide degradation product and a strong mediator of cell metabolism has a great significance in regular erythrocyte shape keeping (Muller et al, 1986). We observed that Ptd Ins-4-P and Ptd Ins-4,5-P2 levels were raised almost threefold in erythrocyte membranes of women with ovarian cancer compared with control subjects.…”

Section: Discussionsupporting

confidence: 58%

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The biochemical modification of the erythrocyte membranes from women with ovarian cancer

Kopczyński

Kuźniak²,

Thielemann³

et al. 1998

Br J Cancer

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Summary The aim of our work was quantitative evaluation of the protein and phospholipid fractions of mature erythrocyte membranes separated from women with ovarian cancer. Blood was sampled from 30 women with ovarian cancer, aged 24-79 years, in the third stage of clinical progression of the disease. Phospholipids were separated from membranes by MOller's acidic extraction method and analysed in thinlayer two-dimensional chromatography. On the silica gel plates nine fractions of phospholipids were separated: sphingomyelin (SPH), phosphatidylethanolamine (PE), phosphatidlyserine (PS), phosphatidylcholine (PC), lysophosphatidylcholine (LPC), phosphatidic acid (PA), phosphatidylinositol (Ptd Ins), phosphatidylinositol-4-phosphate (Ptd Ins-4-P), phosphatidylinositol-4,5-diphosphate (Ptd Ins-4,5-P2). The activity of phospholipase C in erythrocyte membranes was determined by Akhrem's spectrophotometric method. Membrane proteins were separated by polyacrylamide gel electrophoresis, SDS-PAGE. It was shown that PS, SPH, LPC and PA fractions were significantly diminished. The concentration of Ptd Ins-4-P and Ptd Ins-4,5-P2 was significantly increased with simultaneous reduction in Ptd Ins level. The inhibition of phospholipase C reached 80%. The quantitative protein evaluation showed a statistically significant decrease in spectrin and a significant increase in 4.1 protein. The quantitative changes, observed in phospholipid and protein fractions, led to the restructuring of the erythrocyte membrane cytoskeleton, which may be connected to increased susceptibility to haemolysis of red blood cells.Keywords: ovarian cancer; red blood cell; phospholipid metabolism; phospholipase C; red blood cell membrane proteins Ovarian cancer is a common pathology among women over 50 years of age. Up to 2% of the female population will develop ovarian cancer during their lifetime. Cancer exerts a multidirectional influence on the human organism, leading to systemic disturbances. Mature red blood cells are a good experimental model and an easily obtainable material to study these changes. Although erythrocyte does not contain subcellular organelles it is a highly autonomic and specialized cell. That is why many biochemical transformations, difficult to evaluate in other experimental cell studies, are easily observed in erythrocytes. However, little attention has been directed to the effects of cancer on the structure of erythrocyte membrane. Lipids and proteins (Byers et al, 1985;Derick et al, 1992) are the fundamental elements of erythrocyte membrane skeleton. Phospholipids as a basic component of erythrocyte membrane determine its shape and structure and also the influence of external factors on intracellular metabolism (Ferrel et al, 1984;Schwartz et al, 1985;Smith, 1987). Erythrocyte membrane phospholipids are asymmetrically distributed in the two halves of the membrane bilayer. The choline-containing phospholipids phosphatidylcholine and sphingomyelin (PC and SPH) are present mainly in the outer monolayer, whereas the aminophospholipids...

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Section: Discussionsupporting

confidence: 58%

“…Phospholipids were isolated from membranes by the acidic extraction method (Muller et al, 1986). In two-dimensional thin-layer chromatography on the plates covered with silica gel, nine fractions of phospholipids were separated (Broekhuyse, 1969).…”

Section: Phospholipids Isolationmentioning

confidence: 99%

The biochemical modification of the erythrocyte membranes from women with ovarian cancer

Kopczyński

Kuźniak²,

Thielemann³

et al. 1998

Br J Cancer

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“…Our first step was to identify the lipid precursors PtdInsP and PtdInsP 2 and to investigate whether their turnover characteristics are typical of signal precursors. For animals and the green algae Chlamydomonas and Dunaliella, turnover studies have shown that polyphosphoinositides are synthesized and broken down much faster than structural lipids (Mfiller et al 1986;Palmer et al 1986;Einspahr et al 1988;Brederoo et al 1991). Here, evidence for the rapid turnover of polyphosphoinositides in a higherplant system is presented for the first time.…”

Section: Introductionmentioning

confidence: 75%

Rapid turnover of polyphosphoinositides in carnation flower petals

Munnik¹,

Musgrave

Vrije³

1994

Planta

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Abstract. Carnation (Dianthus caryophyllus L. cv. White Sim) petal discs were radiolabelled with [32p]orthophosphate and the lipids were extracted and analysed by thin-layer chromatography and autoradiography. Phospholipids were identified by co-migration with standards using thin-layer chromatography with different solvent systems. Results showed that [32p]orthophosphate was rapidly incorporated into the minor lipids phosphatidic acid (PtdOH), phosphatidylinositol monophosphate (PtdInsP) and phosphatidylinositol bisphosphate (PtdInsP2), and relatively slowly into the structural lipids phosphatidylcholine, -ethanolamine, -glycerol and -inositol. Pulse-chase experiments revealed that the label was rapidly lost from PtdOH, PtdInsP and PtdInsP2 while the structural lipids remained radiolabelled. The amount of PtdInsP and PtdInsP2 was found to constitute 0.45% and 0.013%, respectively, of the total phospholipids, on a molar basis. Together these results show that the turnover of the chemically low-abundant polyphosphoinositides is relatively high compared with the major structural phospholipids. Phosphatidylinositol monophosphate was further characterized by showing that it incorporates myo-[3H]inositol and that its major fatty-acid constituents are palmitic acid and linoleic acid. Furthermore, we present evidence for the presence of both phosphatidylinositol 3-phosphate and phosphatidylinositol 4-phosphate isomers. The significance of these results is discussed with respect to plant phosphoinositide signal transduction.

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“…This may be related to protein phosphorylation powered by ATP that is one of the physiological processes that control membrane stability. One possible candidate is the phosphorylation of the phosphoinositides (PI) because it consumes more ATP than the combined phosphorylation of all the membrane proteins (23). PIP 2 , phosphorylated from PI by ATP, is thought to play an important role in modulating the binding of the lipid bilayer to the cytoskeleton by altering the protein interactions that comprise the junctional complex at spectrin tetramer ends (24).…”

Section: Atp Results In Non-equilibrium Dynamics For Membrane Fluctuamentioning

confidence: 99%

Metabolic remodeling of the human red blood cell membrane

Park

Best²,

Auth

et al. 2010

Proc. Natl. Acad. Sci. U.S.A.

366

362

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The remarkable deformability of the human red blood cell (RBC) results from the coupled dynamic response of the phospholipid bilayer and the spectrin molecular network. Here we present quantitative connections between spectrin morphology and membrane fluctuations of human RBCs by using dynamic full-field laser interferometry techniques. We present conclusive evidence that the presence of adenosine 5′-triphosphate (ATP) facilitates nonequilibrium dynamic fluctuations in the RBC membrane that are highly correlated with the biconcave shape of RBCs. Spatial analysis of the fluctuations reveals that these non-equilibrium membrane vibrations are enhanced at the scale of spectrin mesh size. Our results indicate that the dynamic remodeling of the coupled membranes powered by ATP results in non-equilibrium membrane fluctuations manifesting from both metabolic and thermal energies and also maintains the biconcave shape of RBCs.ATP | imaging technique | membrane fluctuation | RBC | spectrin A s they travel through small blood vessels and organs, RBCs undergo repeated severe deformation. The coupling and interactions between the phospholipid bilayer and the spectrin network govern the deformability of RBCs (1). The fluid-like lipid bilayer is coupled to the two-dimensional spectrin network that comprises an approximately hexagonal lattice via protein junctional complexes. The RBC membrane is remarkably soft and elastic, and thus exhibits fluctuations with amplitudes of the order of tens of nanometers. The dynamics of the RBC membrane is strongly related to the membrane structure and mechanical properties and has been explored extensively (2-6). However, experimental results available to date on RBC membrane fluctuations have provided only limited information on select regions of the cell membrane with limited spatial and/or temporal resolution (7-9). No full-field measurements of membrane fluctuations in the entire RBC arising in response to well-controlled metabolic activity have been made so far and, consequently, different techniques have led to different interpretations of the mechanistic origins of dynamic RBC membrane fluctuations with and without metabolic activity (7-9).The RBC membrane is not a static but a metabolically regulated active structure. It is known that biochemical energy controls its static and dynamic characteristics. The presence of ATP is not only crucial in maintaining the biconcave shape of the RBC membrane (10), but was also shown to increase the dynamic membrane fluctuations (7, 9). However, the regulatory mechanism of ATP in RBC membranes still remains elusive. Furthermore, these static and dynamic effects of ATP on RBC membrane fluctuations have hitherto been regarded as separate phenomena and have never been explored simultaneously.Here, we present dynamic, full-field, and quantitative measurements of ATP effects on RBC membrane morphology and fluctuations. We show that in the presence of ATP, the RBC membrane fluctuations have a non-equilibrium, metabolic component in addition to a thermal o...

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Turnover of phosphomonoester groups and compartmentation of polyphosphoinositides in human erythrocytes

Cited by 53 publications

References 26 publications

The biochemical modification of the erythrocyte membranes from women with ovarian cancer

The biochemical modification of the erythrocyte membranes from women with ovarian cancer

Rapid turnover of polyphosphoinositides in carnation flower petals

Metabolic remodeling of the human red blood cell membrane

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