The Natural Phototransforming Photochrome Membrane Protein Bacteriorhodopsin From Purple Membranes of Halobacterium Halobacterium Halobium

Mosin, Oleg; Ignatov, Ignat

doi:10.13187/ejnr.2014.1.43

Cited by 9 publications

(15 citation statements)

References 6 publications

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“…Therefore, in the case of the strain of obligate methylotrophic bacteria M. flagellatum the inclusion of 13C into amino acid molecules was carried out in one step by growing the bacteria on minimal M9 media containing 1% of [13C]methanol as a source of carbon-13 isotope. As another model system for inclusion of isotopic label into the protein molecules used a transmembrane protein bacteriorhodopsin [21] synthesized in the purple membrane of extreme photo-organotrophic halobacterium Halobacterium halobium ET 1001. The selection for this purpose of bacteriorhodopsin functioning in halobacteria cells as the ATP-dependent translocase was dictated by the ability to use it to study of the functioning of membrane proteins in vivo in deuterium isotope enriched media.…”

Section: Resultsmentioning

confidence: 99%

The Development of Biosynthesis of 2Н- and 13С-labeled Amino acids and Proteins with Various Levels of Isotopic Enrichment Using Bacterial Objects

Mosin¹,

Ignatov²,

Складнев³

et al. 2015

European Reviews of Chemical Research

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By the method of microbiological synthesis were obtained and analyzed by electron impact mass-spectrometry 2H, 13C-labeled amino acids of a facultative methylotrophic bacterium Brevibacterium methylicum and an obligate methylotrophic bacterium Methylobacillus flagellatum and 2H, 13C-labeled amino acids of the total protein of biomass obtained on media containing as a source of stable isotopes [2H]methanol, [13C]methanol and 2H2O. It was also performed the incorporation of L- [2,3,4,5,6-2Н]phenylalanine, L-[3,5-2Н]tyrosine and L- [2,4,5,6,7-2Н]tryptopan into the membrane integral protein bacteriorhodopsin synthesised in purple membranes of photoorganotrophic halobacterium Halobacterium halobium ET 1001. For mass-spectrometric analysis the multicomponential mixtures of 2H-and 13C-labeled amino acids, derived from cultural media and protein hydrolysates after hydrolysis in 6 M 2HСl (3% phenol) and 2 M Ва(OH)2 were modified into N-benzyloxycarbonyl-derivatives of amino acids as well as into methyl esters of N-5-(dimethylamino)naphthalene-1-sulfonyl chloride (dansyl) derivatives of [2H, 13С]amino acids, European Reviews of Chemical Research, 2015, Vol.(5), Is. 3 145 which were preparative separated using a method of reverse-phase HCLP. Biosynthetically obtained 2H-and 13C-labeled amino acids represented the mixtures differing in quantities of isotopes incorporated into molecule. The levels of 2H and 13С enrichment of secreted amino acids and amino acid resigues of protein were found to vary from 20,0 atom.% to L-leucine/isoleucine up to 97,5 atom.% for L-alanine depending on concentration of 2H-and 13C-labelled substrates.Keywords: stable isotopes, methylotrophic bacteria, halobacteria, isotope labeled amino acids, bacteriorhodopsin IntroductionThe enrichment of molecules by stable isotopes (2Н, 13С, 15N, 18O) is an important tool for a variety of biochemical and metabolic studies with amino acids and other biologically active substances (BAS) [1]. The preferential usage of stable isotopes as compared to their counterparts are stipulated by the lack of radioactive radiation hazards and determination of the localization of the label in the molecule by high resolution techniques, including NMR [2], IR spectroscopy [3], and mass spectrometry [4]. The development of these methods for the detection of stable isotopes in biological probes in recent years has significantly increased the efficiency of biological research, as well as studies of the structure and mechanism of action of cellular BAS at the molecular level. In particular, 2H-and 13C-labeled amino acids are used for studying of the spatial structure and conformational changes of proteins, the interaction of protein molecules and in chemical syntheses of some isotope-labeled compounds based on them [5]. For example, isotopically labeled Lphenylalanine is used in the synthesis of peptide hormones and neurotransmitters [6].An important factor in studies using labeled amino acids is their accessibility. 2H and 13C-labeled amino acids can be prepared using ...

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Section: Resultsmentioning

confidence: 99%

The Development of Biosynthesis of 2Н- and 13С-labeled Amino acids and Proteins with Various Levels of Isotopic Enrichment Using Bacterial Objects

Mosin¹,

Ignatov²,

Складнев³

et al. 2015

European Reviews of Chemical Research

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“…The cell membrane of halophiles also contains two sensory rhodopsins, which provide positive and negative phototaxis in cells [17]. These proteins absorb different wavelengths of light, causing a cascade of signals that eventually control the flagella of halobacteria.…”

Section: Resultsmentioning

confidence: 99%

“…Bacterial growth was measured by optical density of the cell suspension at λ=620 nm on a spectrophotometer. As is shown in Figure 6 under optimal growing conditions (incubation period 45 days, temperature t = +35 0 C, illumination with monochromatic light at λ = 560 nm) in cells is synthesized the purple carotenoid pigment, characterized as BR by the spectral ratio of protein and chromophore fragments D280/D568 = 1,5:1,0 in the molecule [37]. The subsequent isolation of BR from the PM fraction is carried out by a combination of physical, chemical and enzymatic methods [38].…”

Section: Biosynthesis Of Brmentioning

confidence: 98%

“…An interesting feature of the metabolism of halobacteria is at that the presence of oxygen and the organic compound (amino acids, peptones), which can be used as growth substrates and sources of energy, halophiles can grow in the dark, by switching on a heterotrophic photosynthetic metabolism [26]. However, with a lack or even in the complete absence of oxygen and under the bright light in the cell membrane of halobacteria is synthesized BR, allowing them to use solar energy for growth and ATP synthesis.…”

Section: The Mechanism Of Functioning Of Brmentioning

confidence: 99%

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Metabolism and Physiology of Halophiles

2015

Russian Journal of Biological Research

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Halophiles (lat. "salt-loving") is the taxonomic group of extreme aerobic obligate Gramnegative microorganisms that live in conditions of high salinity -in the seas, salt lakes, saline soils etc. These microorganisms are known to reddish patina on products, preserved with using large quantities of salt (NaCl). Halophiles were isolated for the first time at the beginning of the XX century from the marine flora estuary mud, but their systematic study was started only at the end of the second decade of the XX century. The internal environment of the human body is not suitable for existence of halobacteria, since none of them are known to have pathogenic forms. Halobacteria have great practical potential for using in molecular bioelectronics and bionanotechnology due to their unique ability to convert the energy of sunlight into electrochemical energy of protons H + due to the presence in their cells a special photo transforming retinal containing integral protein -bacteriorhodopsin, the mechanism of action of which has been currently studied in detail. The paper describes the characteristics of the metabolism and physiology of halophilic bacteria, as well as a method of biosynthesis and preparation of bacteriorhodopsin from purple membranes of cells of the extreme photoorganotrophic halobacterium Halobacterium halobium.

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“…Optical characteristics of BR vary depending on the method of preparation of PM embedded onto the polymer matrix. An interesting feature of the metabolism of halobacteria is at that the presence of oxygen and the organic compound (amino acids, peptones), which can be used as growth substrates and sources of energy, halophiles can grow in the dark, by switching on a heterotrophic photosynthetic metabolism [26]. However, with a lack or even in the complete absence of oxygen and under the bright light in the cell membrane of halobacteria is synthesized BR, allowing them to use solar energy for growth and ATP synthesis.…”

Section: Resultsmentioning

confidence: 99%

Metabolism, Physiology and Biotechnological Applications of Halobacteria

Mosin¹,

Ignatov²

2015

European Journal of Molecular Biotechnology

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Halophiles (lat. "salt-loving") is the taxonomic group of extreme aerobic obligate Gramnegative microorganisms that live in conditions of high salinity -in the seas, salt lakes, saline soils etc. These microorganisms are known to reddish patina on products, preserved with using large quantities of salt (NaCl). Halophiles were isolated for the first time at the beginning of the XX century from the marine flora estuary mud, but their systematic study was started only at the end of the second decade of the XX century. The internal environment of the human body is not suitable for existence of halobacteria, since none of them are known to have pathogenic forms. Halobacteria have great practical potential for using in molecular bioelectronics and bionanotechnology due to their unique ability to convert the energy of sunlight into electrochemical energy of protons H + due to the presence in their cells a special photo transforming retinal containing integral protein -bacteriorhodopsin, the mechanism of action of which has been currently studied in detail. This article describes the characteristics of the metabolism and physiology of halophilic bacteria, as well as a method of biosynthesis and preparation of bacteriorhodopsin from purple membranes of cells of the extreme photoorganotrophic halobacterium Halobacterium halobium.

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The Natural Phototransforming Photochrome Membrane Protein Bacteriorhodopsin From Purple Membranes of Halobacterium Halobacterium Halobium

Cited by 9 publications

References 6 publications

The Development of Biosynthesis of 2Н- and 13С-labeled Amino acids and Proteins with Various Levels of Isotopic Enrichment Using Bacterial Objects

The Development of Biosynthesis of 2Н- and 13С-labeled Amino acids and Proteins with Various Levels of Isotopic Enrichment Using Bacterial Objects

Metabolism and Physiology of Halophiles

Metabolism, Physiology and Biotechnological Applications of Halobacteria

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