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Surface characteristics of phosphatidylglycerol phosphate from the extreme halophile Halobacterium cutirubrum compared with those of its deoxy analogue, at the air/water interface.

ABSTRACT: The relationship between area per molecule and surface pressure of monolayers of phosphatidylglycerol phosphate from extreme halophile Halobacterium cutrirubrum and its deoxy analogue, deoxyphosphatidylglycerol phosphate, spread at an air/water interface was examined. The effect of ionization of the primary and secondary acidic functions of the phosphate groups of the two lipids on surface characteristics of compression isotherms was determined by spreading monolayers on subphases with pH values ranging from below the apparent pKa of the primary ionization (pH 0) to greater than that of secondary ionization (pH 10.9). The limiting molecular area increases with decreasing pH below 2. Ionization of the primary phosphate functions of both phospholipids (with bulk pK1 values close to 4) is associated with a marked expansion of the films, as judged by values of limiting molecular area. Ionization of the secondary phosphate functions causes further expansion of the films, with the apparent pK2 of deoxyphosphatidylglycerol phosphate slightly less than that indicated for phosphatidylglycerol phosphate. Values of surface-compressibility modulus calculated from the surface characteristics of the phosphatidylglcerol phosphate monolayers showed that films spread on subphases with a pH of about the apparent pK1 of the primary phosphate functions were the least compressible. Increasing or decreasing subphase pH caused an increase in compressibility; this effect on compressibility was much less with monolayers of deoxyphosphatidylglycerol phosphate at high pH. The effect of inorganic counter-ions on monolayer characteristics of phosphatidylglycerol phosphate was examined by using subphases of NaCl concentrations varying from 0.01 to 1 M. The limiting molecular area was found to increase exponentially with respect to the subphase NaCl concentration.

SUBMITTER: Quinn PJ

PROVIDER: S-EPMC1138836 | biostudies-other | 1989 Jul

REPOSITORIES: biostudies-other

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Project description:Halobacterium salinarum is a bioenergetically flexible, halophilic microorganism that can generate energy by respiration, photosynthesis, and the fermentation of arginine. In a previous study, using a genome-scale metabolic model, we have shown that the archaeon unexpectedly degrades essential amino acids under aerobic conditions, a behavior that can lead to the termination of growth earlier than necessary. Here, we further integratively investigate energy generation, nutrient utilization, and biomass production using an extended methodology that accounts for dynamically changing transport patterns, including those that arise from interactions among the supplied metabolites. Moreover, we widen the scope of our analysis to include phototrophic conditions to explore the interplay between different bioenergetic modes. Surprisingly, we found that cells also degrade essential amino acids even during phototropy, when energy should already be abundant. We also found that under both conditions considerable amounts of nutrients that were taken up were neither incorporated into the biomass nor used as respiratory substrates, implying the considerable production and accumulation of several metabolites in the medium. Some of these are likely the products of forms of overflow metabolism. In addition, our results also show that arginine fermentation, contrary to what is typically assumed, occurs simultaneously with respiration and photosynthesis and can contribute energy in levels that are comparable to the primary bioenergetic modes, if not more. These findings portray a picture that the organism takes an approach toward growth that favors the here and now, even at the cost of longer-term concerns. We believe that the seemingly "greedy" behavior exhibited actually consists of adaptations by the organism to its natural environments, where nutrients are not only irregularly available but may altogether be absent for extended periods that may span several years. Such a setting probably predisposed the cells to grow as much as possible when the conditions become favorable.

Project description:BACKGROUND:Halobacterium sp. NRC-1 is an extremely halophilic archaeon and has adapted to optimal growth under conditions of extremely high salinity. Its proteome is highly acidic with a median pI of 4.9, a unique characteristic which helps the organism to adapt high saline environment. In the natural growth environment, Halobacterium NRC-1 encounters a number of stressful conditions including high temperature and intense solar radiation, oxidative and cold stress. Heat shock proteins and chaperones play indispensable roles in an organism's survival under many stress conditions. The aim of this study was to develop an improved method of 2-D gel electrophoresis with enhanced resolution of the acidic proteome, and to identify proteins with diverse cellular functions using in-gel digestion and LC-MS/MS and MALDI-TOF approach. RESULTS:A modified 2-D gel electrophoretic procedure, employing IPG strips in the range of pH 3-6, enabled improved separation of acidic proteins relative to previous techniques. Combining experimental data from 2-D gel electrophoresis with available genomic information, allowed the identification of at least 30 cellular proteins involved in many cellular functions: stress response and protein folding (CctB, PpiA, DpsA, and MsrA), DNA replication and repair (DNA polymerase A alpha subunit, Orc4/CDC6, and UvrC), transcriptional regulation (Trh5 and ElfA), translation (ribosomal proteins Rps27ae and Rphs6 of the 30 S ribosomal subunit; Rpl31eand Rpl18e of the 50 S ribosomal subunit), transport (YufN), chemotaxis (CheC2), and housekeeping (ThiC, ThiD, FumC, ImD2, GapB, TpiA, and PurE). In addition, four gene products with undetermined function were also identified: Vng1807H, Vng0683C, Vng1300H, and Vng6254. To study the heat shock response of Halobacterium NRC-1, growth conditions for heat shock were determined and the proteomic profiles under normal (42 degrees C), and heat shock (49 degrees C) conditions, were compared. Using a differential proteomic approach in combination with available genomic information, bioinformatic analysis revealed five putative heat shock proteins that were upregulated in cells subjected to heat stress at 49 degrees C, namely DnaJ, GrpE, sHsp-1, Hsp-5 and sHsp-2. CONCLUSION:The modified 2-D gel electrophoresis markedly enhanced the resolution of the extremely acidic proteome of Halobacterium NRC-1. Constitutive expression of stress proteins and chaperones help the organism to adapt and survive under extreme salinity and other stress conditions. The upregulated expression pattern of putative chaperones DnaJ, GrpE, sHsp-1, Hsp-5 and sHsp-2 under elevated temperature clearly suggests that Halobacterium NRC-1 has a sophisticated defense mechanism to survive in extreme environments.

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Surface characteristics of phosphatidylglycerol phosphate from the extreme halophile Halobacterium cutirubrum compared with those of its deoxy analogue, at the air/water interface.

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