• Volume 15, Issue 3

      September 1990,   pages  117-238

    • Nuclear magnetic resonance and thermal studies of drug doped dipalmitoyl phosphatidyl choline-H2O systems

      K Usha Deniz P S Parvathanathan Geeta Datta C L Khetrapal K V Ramanathan N Suryaprakash S Raghotama

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      The influence of the sulfone drugs, diamino diphenyl sulfone and diamino monophenyl sulfone on the phase transitions and dynamics of dipalmitoyl phosphatidyl choline-H2O/D2O vesicles have been investigated using differential scanning calorimetry and nuclear magnetic resonance. Our results show that diamino diphenyl sulfone interacts quite strongly with the headgroups of dipalmitoyl phosphatidyl choline whereas the diamino monophenyl sulfone-dipalmitoyl phosphatidyl choline interaction is quite weak. This is attributed to the difference in the structure and hydrophobic character of the two drugs.

    • Magnetic resonance methods for studying intact spermatozoa

      Ratna S Phadke Sudha Srivastava Girjesh Govil

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      Motility is used as a routine parameter for assessing spermatozoa activity. The quality rating techniques adopted are based on electron or optical microscopy. However, these methods depend on gross structural and dynamical features of sperm cells and do not provide information on metabolic activity of intact cells. Lately, biochemical assays have become popular. Such methods are cumbersome and destroy the samples. Magnetic resonance methods offer a non-invasive method for studies on intact sperms. We have investigated respiration, maturation andin vitro capacitation of sperms from human ejaculates and sperms extracted from goat reproductive organ using electron spin resonance spin labelling and [31P] nuclear magnetic resonance methods. These studies clearly establish the advantages of magnetic resonance in studies related to metabolic activity of sperms.

    • Erythrocyte stability under imposed fields

      V Sitaramam

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      Lysis of erythrocytes offers an unique opportunity to probe the fine structure of the bilayer as a function of its state of energization. Critical monitoring of the volumes, ion fluxes and related measures in erythrocytes exposed to a variety of milieu and treatments showed that one can critically distinguish the nature of the prelytic perturbations and the proximate forces actually responsible for the disruption of the membranes among surface charge density, elastic energy etc.

    • Depth profiling in membranes by fluorescence quenching

      Amitabha Chattopadhyay

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      Membrane penetration depth is an important parameter in relation to membrane structure and organization. A methodology has been developed to analyze the membrane penetration depths of fluorescent molecules or groups utilizing differential fluorescence quenching caused by membrane embedded spin-label probes located at different depths. The method involves determination of the parallax in the apparent location of fluorophores, detected when quenching by phospholipids spin-labelled at two different depths is compared. By use of relatively simple algebraic expressions, the method allows calculation of depth in å. This method has been used to determine the location of fluorophores in NBD-labelled lipids and anthroyloxy-labelled fatty acids in model membranes and of the membrane embedded tryptophan residues in the reconstituted nicotinic acetylcholine receptor.

    • Role of nicotinic acid as modulator of liposomal microviscosity

      M Chatterjee R Basu P Nandy

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      Using the fluorescent probe 1,6-diphenyl-1,3,5 hexatriene, we have investigated the effect of nicotinic acid, a derivative of the toxic alkaloid nicotine, on the fluidity profile and activation energy of diffusion in the liposomal system of several lipids. We have also studied how the fluidizing property of nicotinic acid affects the intermediate fluid condition induced by cholesterol in these liposomal systems

    • Fluidity of detergent micelles plays an important role in muscarinic receptor solubilization

      Anu KŎiv Ago Rinken Jaak Järv

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      In order to find a suitable reagent for extracting the muscarinic receptor from rat brain membranes 14 different detergents were tested. Only the plant glycoside digitonin efficiently solubilized the receptor protein in its native form. At the same time microviscosity of detergent micelles was determined by measuring the fluorescence polarization of a hydrophobic fluorescent probe diphenylhexatriene incorporated into the micelles. In the case of digitonin the polarization value was close to the corresponding value obtained for rat brain membrane fragments, while for the other detergents studied it remained considerably lower. The results obtained indicate that the fluidity of detergent micelles may play an important role in retaining the active conformation of the solubilized muscarinic receptor.

    • Peptide induced polymorphism in model membranes

      Ratna S Phadke Sudha Srivastava Girjesh Govil

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      Lipids in biological membranes generally adopt bilayer structures. However, incorporation of peptides may induce alterations in such structures. We have studied the influence of tryptophan, leucine, Trp-Leu, luteinizing hormone releasing hormone and renin inhibitor peptide on lipid organisation in liposomes. It has been observed that the effect is specific to the peptide molecule as-a whole and does not have direct correlation to the constituent amino acids or the conformation of the molecule.

    • Kinetic mechanisms of mitochondrial carriers catalysing exchange reactions

      F E Sluse A Evens C Duyckaerts C M Sluse-Goffart

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      The single-binding site or ping-pong mechanism is widely accepted for exchange reactions, catalysed by mitochondrial carriers.

      However, when the most relevant approach to discriminate between mechanisms,i.e., kinetic study is used, the ping-pong mechanism is eliminated in favour of the sequential or ternary complex mechanism implying two binding sites simultaneously accessible to both internal and external substrates. This is the case for the oxoglutarate carrier, the aspartate/glutamate carrier and there are very strong presumptive evidences for the adenylic carrier.

    • Identification and isolation of ATP transport protein in mycobacillin sensitive Aspergillus niger

      Bhabadeb Chowdhury S K Bose

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      The temperature sensitive release and uptake of ATP through theAspergillus niger G3Br membrane vesicles followed saturation kinetics. Both the processes which occurred in the absence of mycobacillin were greatly enhanced by its presence. Liposomes prepared with antifilipin sterol and lipid showed the release and uptake of ATP in the presence of filipin, but no such uptake and release was seen with antimycobacillin sterol and lipid in the presence of mycobacillin. However the liposomes supplemented withAspergillus niger membranes protein (s) showed the release and uptake of ATP, implicating membrane protein as a carrier in the transport process.

    • pH-Dependent membrane interactions of diphtheria toxin: A genetic approach

      R John Collier

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      A genetic approach is described for exploring the mechanism by which diphtheria toxin undergoes pH-dependent membrane insertion and transfer of its enzymic A fragment into the cytoplasm of mammalian cells. The cloned toxin expressed inEscherichia coli is secreted to the periplasmic space, where it is processed normally and folds into a native structure. When bacteria synthesizing the toxin are exposed to pH 5, they die rapidly. The toxin undergoes a conformational change that is believed to allow it to be inserted into the bacterial inner membrane and form channels, which proves lethal for the cell. The membrane insertion event mimics the process by which the toxin inserts into the endosomal membrane of mammalian cells, leading to release of the enzymic A fragment into the cytoplasm. The observation of pH-dependent bacterial lethality provides the basis for a positive genetic selection method for mutant forms of the toxin that are altered in ability to undergo membrane insertion or pore formation.

    • Iron-regulated membrane proteins and bacterial virulence

      E Griffiths

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      The amount of iron that might be readily available to bacteria in body fluids is extremely small. This iron restricted environment induces phenotypic changes in the metabolism and in the composition of the membrane proteins of bacteria growingin vivo. These changes are now providing a fresh insight into the capabilities of bacteria to multiply in host tissues and are suggesting new possibilities for targetting therapeutic and prophylactic measures.

    • Inhibition of anion transport in the red blood cell membrane by anionic and non-anionic arginine-specific reagents

      Laila Zaki

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      Arginine specific reagents are found to be powerful inhibitors of anion exchange in the red blood cell membrane. Some of these inhibitors such as cyclohexandione, phenylglyoxal and 2, 3-butandione are found to produce their inhibition by interacting covalently with band 3. In contrast to the action of these compounds, the inhibition caused by the phenylglyoxal derivative 4-hydroxy-3-nitrophenyl-glyoxal has been found to be completly reversible. In extending the studies on the mode of action of these compounds on sulfate exchange and to get some more information about their binding site, the degree of inhibition caused by different phenylglyoxal derivatives which have a similar core but differ in their substituent groups have been compared. The interaction between the binding sites of these compounds and other anion transport inhibitors have also been studied.

    • Functional and pathological significance of phospholipid asymmetry in erythrocyte membranes

      Robert A Schlegel Susan Kemper Patrick Williamson

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      The normal asymmetric distribution of phospholipids in the plasma membrane is perturbed in erythrocytes from patients with chronic myelogenous leukemia. Since experimentally-produced lipid-symmetric erythrocytes are more interactive with cells of the reticuloendothelial system than are their lipid-asymmetric counterparts, the biological recognition of chronic myelogenous leukemia erythrocytes by the reticuloendothelial system was examined. With one exception, all erythrocyte samples from patients with chronic/benign chronic myelogenous leukemia were more adherent to endothelial cells and more readily phagocytosed by macrophagesin vitro than were normal erythrocytes. Thus, these naturally occurring pathological erythrocytes display the same dysfunctional intercellular interactions as the laboratory models.

    • Calcium and magnesium induced changes in the relative fluidity of phosphatidylcholine liposomes

      R K Mishra Gauri S Singhal

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      The effect of Ca2+ and Mg2+ on relative fluidity of phosphatidylcholine liposomes was studied by measuring the degree of chlorophyll fluorescence polarization. An increase in the degree of fluorescence polarization was observed on incubation of liposomes with different concentrations of Ca2+ or Mg2+. The results have been interpreted on the basis of increase in the size of liposomes which could be brought about by calcium or magnesium induced fusion of small unilamellar liposomes to form larger vesicles. Fusion of liposomes has also been confirmed by the experiments on efficiency of energy transfer from chlorophyll b to chlorophyll a, and transmission electron microscopy of liposomes before and after incubation with Ca2+ and Mg2+.

    • Expression of 5-amino levulinic acid induced photodynamic damage to the thylakoid membranes in dark sensitized by brief pre-illumination

      N Chakraborty B C Tripathy

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      The 5-amino levulinic acid treated cucumber (Cucumis sativus L., CV. Pointsette) plants upon exposure to light (≃30,000 lux) wilted within 6 h and died after 36 h due to photodynamic reactions. Thylakoid membranes, the site of accumulation of porphyrins, were damaged due to photodynamic reactions leading to the inhibition of membrane linked functions of photosystem II, photosystem I and the whole chain electron transport. Photosystem II was more susceptible to photodynamic damage than photosystem I. The exogenous electron donors Mn2+, diphenyl carbazide and NH2OH failed to donate electrons to photosystem II suggesting that the damage has taken place close to P680. The 5-amino levulinic acid treated plants exposed to 30 min of light did not show any damage to the thylakoid membranes. However, when the above plants were transferred to dark for 12 h there was substantial damage to the thylakoid membrane system.

    • Polyvanadate acts at the level of plasma membranes through α-adrenergic receptor and affects cellular calcium distribution and some oxidation activities

      T Ramasarma Sharada Gullapalli Vidya Shivaswamy C K Ramakrishna Kurup

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      The activities of calcium-stimulated respiration, calcium uptake, α-glycero-phosphate dehydrogenase and rates of oxidation in state 3 and of H2O2 generation, were found to increase and that of pyruvate dehydrogenase decrease in mitochondria isolated from livers of rats administered intraperitoneally or perfused with polyvanadate. Phenoxybenzamine, an antagonist of α-adrenergic receptor, effectively prevented these changes. It was also found that perfusion of the liver with polyvanadate reproduced one of the best characterized events of α-adrenergic activation-stimulation of protein kinase C in plasma membrane accompanied by its decrease in cytosol. These experiments indicate for the first time the α-adrenergic mimetic action of polyvanadate.

    • Membrane lipid peroxidation by ultrasound: Mechanism and implications

      A K Jana S Agarwal S N Chatterjee

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      Ultrasonic radiation produced a dose-dependent linear increase in lipid peroxidation in the liposomes membrane as reflected in the measurement of conjugated dienes, lipid hydroperoxides and malondialdehydes. Ultrasound induced malondialdehyde production could not be inhibited by any significant degree by superoxide dismutase or histidine or dimethyl furan but was very significantly inhibited by butylated hydroxytoluene, cholesterol, sodium benzoate, dimethyl sulphoxide, sodium formate and EDTA. The scavenger studies indicated the functional role of hydroxyl radicals in the initiation of ultrasound induced lipid peroxidation.

    • Dephosphorylation of cell-surface phosphoproteins of goat spermatozoa

      M Barua G C Majumder

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      Multiple ecto-phosphoproteins of the goat cauda-epididymal intact spermatozoa have been shown to undergo dephosphorylationin vitro by endogenous phosphoprotein phosphatase(s) located on the sperm outer surface. The major ecto-phosphoproteins that are dephosphorylated have molecular masses of 27, 40, 70, 116 and 205 kDa. The cell surface dephosphorylation reaction is not dependent on bivalent metal ions. Mg2+ (5 mM), Mn2+ (5 mM), orthovanadate (200ΜM) and cAMP (5 ΜM) have no effect on this surface reaction whereas it is inhibited nearly 50% by Co2+ or Zn2+ (1 mM). Spermidine (5 mM), or Ca2+ (1mM) inhibited to a small extent (approx. 25%) the cell surface dephosphorylation of proteins.

    • Activities of myelin bound cytidine 5′-diphosphate-choline 1, 2 diacyl-glycerol choline phosphotransferase and uridine 5′-diphosphate-galactose-ceramide galactosyltransferase under restricted food intake

      S Padmini P Srinivasarao

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      Activities of cytidine 5′-diphosphate-choline glycerol choline phosphotransferase and uridine 5′-diphosphate galactose-ceramide galactosyltransferase were determined in isolated myelin in different brain regions of control, and rats with restricted food intake. Kinetic experiments indicated an increase inKm value of phosphocholinetransferase in brain stem of undernourished rats, without significant change in the specific activity of this enzyme. Stimulation of this myelin bound enzyme activity was also evident in the animals when myelin was treated with the detergent: Tween CF. 54. Though specific activities of galactosyl transferase in myelin of undernourished rats were significantly diminished, theKm of this enzyme was unaltered. These studies point to an adverse effect of early nutritional stress on the activities of enzymes bound to myelin membrane which has hitherto been considered metabolically inert.

    • Desmosome and intermediate filament assembly during differentiation and stratification of epithelial cells

      Asima Lahiri Majumder Charles F Shuler

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      In the present investigation the sequential expression and organization of keratin intermediate filament proteins were studied in the developing rat palatal epithelia starting from early gestation period to the adult. The distribution and organization of keratin proteins were correlated with the formation and elaboration of desmosornes during differentiation and stratification of the epithelia.

    • Coupling of proteins to liposomes and their role in understanding delayed type of hypersensitivity in human and mice

      U Sengupta Sudhir Sinha V Chaturvedi R B Narayanan Sreevatsa C M Gupta

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      Liposome-coupled lepromin was found to elicit a 3-week skin reaction in leprosy patients similar to that elicited by wholeMycobacterium leprae. The present study suggests that the presentation of antigens in a specific orientation is necessary for evoking delayed type hypersensitivity response in humans.

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