Sample records for alanine-alpha

  1. Beta-alanine/alpha-ketoglutarate aminotransferase for 3-hydroxypropionic acid production

    Jessen, Holly Jean; Liao, Hans H; Gort, Steven John; Selifonova, Olga V


    The present disclosure provides novel beta-alanine/alpha ketoglutarate aminotransferase nucleic acid and protein sequences having increased biological activity. Also provided are cells containing such enzymes, as well as methods of their use, for example to produce malonyl semialdehyde and downstream products thereof, such as 3-hydroxypropionic acid and derivatives thereof.

  2. Beta-alanine/alpha-ketoglutarate aminotransferase for 3-hydroxypropionic acid production

    Jessen, Holly Jean (Chanhassen, MN); Liao, Hans H. (Eden Prairie, MN); Gort, Steven John (Apple Valley, MN); Selifonova, Olga V. (Plymouth, MN)


    The present disclosure provides novel beta-alanine/alpha ketoglutarate aminotransferase nucleic acid and protein sequences having increased biological activity. Also provided are cells containing such enzymes, as well as methods of their use, for example to produce malonyl semialdehyde and downstream products thereof, such as 3-hydroxypropionic acid and derivatives thereof.

  3. Identification of a mutation affecting an alanine-alpha-ketoisovalerate transaminase activity in Escherichia coli K-12.

    Falkinham, J O


    A mutation affecting alanine-alpha-ketoisovalerate transaminase activity has been shown to be cotransducible with ilv gene cluster. The transaminase deficiency results in conditional isoleucine auxotrophy in the presence of alanine. PMID:396446

  4. Parametrization, molecular dynamics simulation and calculation of electron spin resonance spectra of a nitroxide spin label on a poly-alanine alpha helix

    Sezer, Deniz; Freed, Jack H.; Roux, Benoît


    The nitroxide spin label 1-oxyl-2,2,5,5-tetramethylpyrroline-3-methyl-methanethiosulfonate (MTSSL), commonly used in site-directed spin labeling of proteins, is studied with molecular dynamics (MD) simulations. After developing force field parameters for the nitroxide moiety and the spin label linker, we simulate MTSSL attached to a poly-alanine alpha helix in explicit solvent to elucidate the factors affecting its conformational dynamics. Electron spin resonance spectra at 9 and 250 GHz are ...

  5. Solvated States of Poly-L-alanine alpha-Helix Explored by Raman Optical Activity

    Yamamoto, S.; Furukawa, T.; Bouř, Petr; Ozaki, Y.


    Roč. 118, č. 20 (2014), s. 3655-3662. ISSN 1089-5639 R&D Projects: GA ČR GAP208/11/0105 Grant ostatní: AV ČR(CZ) M200550902 Institutional support: RVO:61388963 Keywords : molecular property tensors * nuclear magnetic resonance * activity spectra Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.693, year: 2014

  6. Non-specific esterases and esterproteases in masticatory muscles from the muscular dystrophic mouse

    Kirkeby, S; Moe, D; Vilmann, H


    With the aid of histochemical and electrophoretic techniques activities for esterase and esterprotease were investigated in the digastric and masseter muscles from normal and dystrophic mice. The substrates used were alpha-naphthyl acetate and N-acetyl-L-alanine alpha-naphthyl ester. According to...

  7. Demonstration of murine pancreas elastase and its interstrain variation by isoelectric focusing.

    von Deimling, O H; Gaa, A; Hameister, H


    Isoelectric focusing between pH 9 and 11 was used for separation of murine pancreas proteases. One of these proteases is characterized by its preference for N-acetyl-L-alanine-alpha-naphthylester as substrate and by its genetic linkage to bt, a coat color marker of chromosome 15. This protease was identified as elastase, and is probably elastase-1 (ELA-1). Because of the simple procedure and the excellent reproducibility of the focusing pattern, ELA-1 is recommended as a useful marker for mouse chromosome 15. PMID:1889396

  8. [Transaminase activity of the cortical layer of the kidney of rats of different ages and sex after administration of hydrocortisone and insulin].

    Poletaeva, K A


    Response of cortical layer of rat kidney to separate and combined administration of hydrocortisone and insulin, as manifested by the activity of aspartate-alpha-ketoglutarate transaminase (Asp-T) and alanine-alpha-ketoglutarate transaminase (Ala-T), varied in males and females of different age. Prolonged administration of insulin to normal preadolescent rats and to adult males and females did not affect the activity of Asp-T and Ala-T in the cortical layer of kidney. During simultaneous prolonged administration of hydrocortisone and insulin to preadolescent male rats, there occurred no increase in the activity of Asp-T induced by administration of hydrocortisone alone. During simultaneous prolonged administration of hydrocortisone and insulin to adult male rats, activity of Asp-T of the cortical layer of kidney remained at the same level at after administration of hydrocortisone alone. PMID:5317624

  9. The isolation and partial characterization of the plasma membrane from Trypanosoma brucei.

    Voorheis, H P; Gale, J S; Owen, M J; Edwards, W


    Whole sheets of plasma membrane, each with their attached flagellum, were purified from Trypanosoma brucei. The method devised for their isolation included a new technique of cell breakage that used a combination of osmotic stress followed by mechanical sheer and avoided the problem of extreme vesiculation as well as the trapping of organelles in cell 'ghosts'. The purified membranes all contained the pellicular microtubular array. The antigenic surface coat was completely released from the plasma membrane during the isolation procedure. The membranes had a very high cholesterol/phospholipid ratio (1.54). A large proportion (42%) of the cellular DNA was recovered in the plasma-membrane fraction unless a step involving deoxyribonuclease treatment, which decreased the DNA content to less than 13%, was included before secrose-density gradient centrifugation. This step also aided the separation of plasma membranes from other cellular components. The ouabain-sensitive Na+ + K+-stimulated adenosine triphosphatase and adenylate cyclase co-purified with the plasma membranes. Although 5'-nucleotidase was thought to be a plasma-membrane component, it was easily detached from the membrane. The purified membranes were essentially free of L-alanine-alpha-oxoglutarate aminotransferase, L-asparte-alpha-oxoglutarate aminotransferase, malate dehydrogenase, oligomycin-sensitive adenosine triphosphatase, glucose 6-phosphatase, Mg2+-stimulated p-nitrophenyl phosphatase and catalase. PMID:486094

  10. Carbon-11 labelled analogs of alanine by the Strecker synthesis

    Prenant, C.; Theobald, A.; Siegel, T.; Joachim, J.; Weber, K.; Haberkorn, U.; Oberdorfer, F. [Deutsches Krebsforschungszentrum, Heidelberg (Germany)


    Derivatives of alanine, {alpha}-[2-sup(11)C]aminoisobutyric acid 1a and {alpha}-(N-methyl)-[2-{sup 11} C]aminoisobutyric acid 1b were prepared for the in-vivo study of amino acid transport phenomena by positron-emission-tomography (PET). Compounds 1a and 1b were obtained by a Zelinski-Stadnikoff variant of the Strecker {alpha}-amino acid synthesis from in-situ formed [{sup 11} C]acetone in presence of sodium cyanide and either ammonium sulfate (for 1a) or methylamine hydrochloride (for 1b). The complete preparation required 50 min from the end of [{sup 11}C]CO{sub 2} production, and delivered 1.2 - 2 GBq of labelled product for application (2.4 -4%); not corrected for decay; related to trapped [{sup C}]CO{sub 2}. The specific activity of the labelled products was 16 to 20 GBq{center_dot}{mu}mol{sup -1}. The radiochemical and chemical purity of the preparations was greater than 98%. (Author).

  11. Human red cell butyrylesterase, and its homologies in thirteen other mammalian species.

    von Deimling, O; de Looze, S


    The selective staining of a single butyrylesterase, following isoelectric focusing of red cell lysates from 14 mammalian species, including man, was achieved using the chromogenic substrate N-acetyl-L-alanine-alpha-naphthyl ester. This procedure optimized the identification of this enzyme, and a close correspondence of its properties was observed in all the species investigated. The inhibition profile, using a range of inhibitors, was identical in all cases. Particularly noteworthy was the activation of the enzyme by p-chloromercuribenzoate at low (2-20 x 10(-6) M) concentrations, but inhibition at higher (greater than 10(-3) M) concentrations. The isoelectric points of all samples fell within a narrow range, pI 4.00-4.43. Descriptions of this activity in different animals, under different designations, could be identified in earlier studies, and the enzyme was uniformly designated "butyrylesterase 1". A tetrameric subunit structure, previously established for the human enzyme, was also demonstrated in the case of mouse butyrylesterase 1. A list of diagnostic characteristics for the enzyme was presented, and it was assumed that the 14 enzymes investigated are orthologous. It was not possible to group butyrylesterase 1 with any other known esterase within the system of enzymes recommended by the IUB, and it was proposed that this enzyme be assigned to a new esterase subclass. PMID:6852820

  12. Relation between track structure and LET effect on free radical formation for ion beam-irradiated alanine dosimeter

    Krushev, V.V.; Koizumi, Hitoshi; Ichikawa, Tsuneki; Yoshida, Hiroshi [Hokkaido Univ., Sapporo (Japan). Faculty of Engineering; Shibata, Hiromi; Tagawa, Seiichi [Tokyo Univ. (Japan). Research Center for Nuclear Science and Technology; Yoshida, Yoichi [Tokyo Univ. (Japan). Faculty of Engineering


    The yield and local concentration of free radicals generated from alanine ({alpha}-aminopropionic acid) by irradiation with 3 MeV H{sup +} and He{sup +} ions were examined by means of electron spin resonance (ESR) and ESR power saturation methods at room temperature. The G-value of the radical formation showed a marked dependence on linear energy transfer (LET) of the ions. The G-value for the H{sup +} ion (average LET: 28 eV/nm) was almost the same as that for {gamma}-irradiation and it was smaller by a factor of 1/4.7 for the He{sup +} ion (average LET: 225eV/nm). Combining the local concentration of the free radicals along the ion tracks with the G-values and the reported ion range, the radius of a track filled with free radicals was estimated to be 4 {approx} 5 nm by assuming a simple rod-shaped track with a constant radius and homogeneous distribution of the free radicals in it. The track radius scarcely depends on the LET within the range examined. The radiation energy deposited in the core region of the ion track was concluded to spread over the rod to generate free radicals. (author).

  13. Hyper-beta-alaninemia associated with beta-aminoaciduria and gamma-aminobutyricaciduaia, somnolence and seizures.

    Scriver, C R; Pueschel, S; Davies, E


    Hyper-beta-alaninemia was found in a somnolent, convulsing infant. Hyper-beta-aminoaciduria (beta-ala, betaAIB and taurine) was also observed, varying directly with plasma beta-alanine concentration. The beta-aminoaciduria is explained by the interaction between beta-alanine and a specific cellular-transport system with preference for beta-amino compounds. Gamma-aminobutyricaciduria was also observed, its excretion being independent of beta-alanine levels. Dietary modifications, pyridoxine, pantothenic acid and antibiotic therapy were not beneficial. Post-mortem tissues had elevated levels of beta-alanine and carnosine; GABA levels in brain were probably elevated for the age of the patient. A proposed block in beta-alanine-alpha-ketoglutarate transaminase would expand the free beta-alanine pool, thus increasing tissue carnosine. beta-Alanine is a central-nervous-system depressant. Associated inhibition of GABA transaminase and displacement of GABA from central-nervous-system binding sites would produce GABAuria and convulsions. PMID:17926374

  14. Photoion mass spectrometry of five amino acids in the 6-22 eV photon energy range

    Jochims, Hans-Werner; Schwell, Martin; Chotin, Jean-Louis; Clemino, Monique; Dulieu, Francois; Baumgaertel, Helmut; Leach, Sydney


    A photoionization mass spectrometry study in the 6-22 eV photon energy region of five amino acids, glycine-h{sub 5} and its -d{sub 5} isotopologue, {alpha}-alanine, {beta}-alanine, {alpha}-aminoisobutyric acid and {alpha}-valine, revealed VUV-induced degradation pathways of these important biological molecules. The fragmentation patterns, ionization energies and ion appearance energies are reported, many for the first time, and are compared with results of electron impact and other studies. Assignment of ion peaks and determination of fragment ion formation channels were assisted by mass spectral data on deuterated isotopologues of the three proteinaceous amino acids studied. Thermochemical data, coupled with the observed ion appearance energies, was also useful in clarifying dissociative photoionization pathways. Ion pair formation appears to occur in certain low energy dissociation processes. Isomeric interconversion between {alpha}-alanine and {beta}-alanine cations does not occur up to 20 eV excitation energy. Some astrophysical implications concerning the prospects for amino acid observation and survival in the interstellar medium and in meteorites are briefly discussed.

  15. Genetics of a tissue esterase polymorphism (Est-6) in the rabbit (Oryctolagus cuniculus).

    van Zutphen, L F; den Bieman, M G; von Deimling, O; Fox, R R


    Genetic analysis of a polymorphic tissue esterase revealed a new locus (Est-6) with two alleles (Est-6a and Est-6b) on linkage group VI of the rabbit. Est-6 is closely linked to the Est-1,2,4 cluster. Esterase of Est-6 is found in many organs, particularly in liver and small intestine, but not in erythrocytes and serum. Est-6 esterase hydrolyzes alpha-naphthyl acetate and butyrate, naphthol AS-D acetate, indoxyl acetate, and butyrate as well as 5-bromoindoxyl acetate, N-acetyl-L-alanine-alpha-naphthyl ester but not 4-methylumbelliferyl acetate and fluorescein diacetate. The enzyme is inhibited by bis-p-nitrophenyl phosphate and eserine but not by p-chloromercuribenzoate. It was classified as a carboxylesterase (EC Based on chromosomal localization, tissue distribution, substrate specificity, inhibitor sensitivity, and range of pI's, rabbit Est-6 is assumed to be homologous with mouse Es-7. PMID:3619880

  16. Acute hepatotoxicity of ethylene and halogenated ethylenes after PCB pretreatment.

    Conolly, R B; Jaeger, R J


    Previous studies from our laboratory have shown that ethylene, vinyl fluoride monomer (VFM), vinyl chloride monomer (VCM), and vinyl bromide monomer (VBM) are all acutely hepatotoxic in rats pretreated with polychlorinated biphenyl (PCB). The time course of hepatic injury development after exposure and several parameters, environmental and chemical, affecting this toxicity were evaluated in the work reported here. Liver injury, as measured by serum alanine-alpha-ketoglutarate transaminase (SAKT) or sorbitol dehydrogenase (SDH), develops progressively over a 24-hr period following a 4-hr inhalation exposure of PCB-pretreated rats to ethylene or VCM. Environmental temperature during exposure to VCM does not affect hepatotoxicity or mortality below 30.3 degrees C. At 33.8 degrees C, however, mortality and SAKT are dramatically increased. Overnight fasting, which depletes hepatic glutathione (GSH) of PCB-pretreated rats before exposure to ethylene or VCM, significantly increases the hepatotoxicity of these compounds as measured by SDH. The combined effects of fasting and of trichloropropane epoxide (TCPE), an inhibitor of epoxide hydrase (EH), were also examined. TCPE treatment of fasted PCB-pretreated rats immediately before exposure was synergistic in increasing the acute toxicity of ethylene and VCM. TCPE increased mortality in fed or fasted rats exposed to VFM, but there was no effect of fasting alone. Both fasting and TCPE increased the sensitivity of PCB-pretreated rats to VBM, but there was not a clearly synergistic effect of fasting plus TCPE. These data suggest that the acute toxicity of these compounds is mediated through epoxide intermediates. PMID:417916

  17. Regulation of transaminase C synthesis in Escherichia coli: conditional leucine auxotrophy.

    McGilvray, D; Umbarger, H E


    The regulation of synthesis of the valine-alanine-alpha-aminobutyrate transaminase (transaminase C) was studied in Escherichia coli mutants lacking the branched-chain amino acid transaminase (transaminase B). An investigation was made of two strains, CU2 and CU2002, each carrying the same transaminase B lesion but exhibiting different growth responses on a medium supplemented with branched-chain amino acids. Both had the absolute isoleucine requirement characteristic of ilvE auxotrophs, but growth of strain CU2 was stimulated by valine, whereas that of strain CU2002 was markedly inhibited by valine. Strain CU2002 behaved like a conditional leucine auxotroph in that the inhibition by valine was reversed by leucine. Results of enzymatic studies showed that synthesis of transaminase C was repressed by valine in strain CU2002 but not in strain CU2. Inhibition by valine in strain CU2002 appears to be the combined effect of repression on transaminase C synthesis and valine-dependent feedback inhibition of alpha-acetohydroxy acid synthase activity, causing alpha-ketoisovalerate (and hence leucine) limitation. The ilvE markers of strains CU2 and CU2002 were each transferred by transduction to a wild-type genetical background. All ilvE recombinants from both crosses resembled strain CU2002 and were inhibited by valine in the presence of isoleucine. Thus, strain CU2 carries an additional lesion that allows it to grow on a medium containing isoleucine plus valine. It is concluded that conditional leucine auxotrophy is characteristic of mutants carrying an ilvE lesion alone. PMID:4616947

  18. Variations in elastaselike esterase activities in human leucocytes during cell maturation.

    Feinstein, G; Janoff, A


    Granules of human peripheral blood leucocytes contain four well-characterized elastase isozymes and one or two slow-moving elastaselike esterases (SE) which have not been as well characterized. SE are capable of hydrolyzing typical elastase synthetic sybstrates such as N-acetyl-dl-alanine-alpha-naphthyl ester (Ac-DL-Ala-1-ONap) and N-t-butyloxycarbonyl-L-alanine-p-nitrophenyl ester (Boc-Ala-ONp), but unlike the highly basic elastase isozymes, SE barely migrate into 13% acrylamide gels during cationic electrophoresis at pH 4.3. Hydrolysis of Ac-DL-Ala-1-ONap by SE requires the presence of Triton in the gel, and hydrolysis of Boc-Ala-ONp by the same enzyme(s) is also enhanced in the presence of the detergent. Triton is not required for these activities, in the case of the elastase isozymes. Diisopropylfluorophosphate (Dip-F) inactivates both SE and the elastase isozymes, whereas Ac-(Ala)2-Pro-AlaCH2Cl (a powerful inactivator of the leucocyte elastase isozymes at 10-4 M concentration) does not inactivate SE at the same concentration. Immunochemical studies revealed antigenic cross-reaction between the rapidly migrating leucocyte elastase isozymes and SE. Two preparations of leucocyte granules from nonleukemic bone marrow cells showed no activity of the rapidly migrating elastase isozymes, but did contain SE activity. SE may be a precursor or zymogen form of the elastase isozymes, present in immature cells and partly retained through later stages of development. PMID:1265076

  19. The cisproline(i - 1)-aromatic(i) interaction: Folding of the Ala-cisPro-Tyr peptide characterized by NMR and theoretical approaches

    Nardi, Frederico; Kemmink, Johan; Sattler, Michael; Wade, Rebecca C. [European Molecular Biology Laboratory (Germany)


    Cisproline(i-1)-aromatic(i) interactions have been detected in several short peptides in aqueous solution by analysis of anomalous chemical shifts measured by {sup 1}H-NMR spectroscopy. This formation of local structure is of importance for protein folding and binding properties. To obtain an atomic-detail characterisation of the cisproline(i-1)-aromatic(i) interaction in terms of structure, energetics and dynamics, we studied the minimal peptide unit, blocked Ala-cisPro-Tyr, using computational and experimental techniques. Structural database analyses and a systematic search revealed two groups of conformations displaying a cisproline(i-1)-aromatic(i) interaction. These conformations were taken as seeds for molecular dynamics simulations in explicit solvent at 278 K. During a total of 33.6 ns of simulation, all the 'folded' conformations and some 'unfolded' states were sampled. {sup 1}H- and {sup 13}C-chemical shifts and {sup 3}J-coupling constants were measured for the Ala-Pro-Tyr peptide. Excellent agreement was found between all the measured and computed NMR properties, showing the good quality of the force field. We find that under the experimental and simulation conditions, the Ala-cisPro-Tyr peptide is folded 90% of the time and displays two types of folded conformation which we denote 'a' and 'b'. The type a conformations are twice as populated as the type b conformations. The former have the tyrosine ring interacting with the alanine {alpha} proton and are enthalpically stabilised. The latter have the aromatic ring interacting with the proline side chain and are entropically stabilised. The combined and complementary use of computational and experimental techniques permitted derivation of a detailed scenario of the 'folding' of this peptide.

  20. Interaction of some hydrophobic amino acids, peptides, and protein with aqueous 3-chloro-1,2-propanediol and 3-chloro-1-propanol: Biophysical studies

    Keswani, Neelam [Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400 076 (India); Kishore, Nand, E-mail: [Department of Chemistry, Indian Institute of Technology Bombay, Powai, Mumbai 400 076 (India)


    Research highlights: Thermodynamic properties of amino acids, peptides and protein determined in solution. The solvents chosen were 3-chloropropan-1-ol and 3-chloropropan-1,2-diol. {yields}The results enabled understanding the interactions quantitatively in these systems affecting the protein stability. Fine details of interactions provided in-depth analysis. - Abstract: The apparent molar volume V{sub 2,{phi},} apparent molar isentropic compressibility K{sub S,2,{phi},} and heat of dilution (q) of aqueous glycine, alanine, {alpha}-amino butyric acid, valine, leucine, diglycine, triglycine, and hen egg white lysozyme have been determined in aqueous solutions of 3-chloropropano-1-ol and 3-chloropropan-1,2-diol solutions at T = 298.15 K. These data have been used to calculate the infinite dilution standard partial molar volume V{sub 2,m}{sup 0}, partial molar isentropic compressibility K{sub S,2,m}{sup 0}, and enthalpy of dilution {Delta}{sub dil}H{sup o} of the amino acids and peptides in aqueous 3-chloropropano-1-ol and 3-chloropropan-1,2-diol, and the standard partial molar quantities of transfer of the amino acids and peptides to the aqueous alcohol and diol solutions. The linear correlation of V{sub 2,m}{sup 0} for a homologous series of amino acids has been utilized to calculate the contribution of the charged end groups (NH{sub 3}{sup +},COO{sup -}), CH{sub 2} group and other alkyl chains of the amino acids to the values of V{sub 2,m}{sup 0}. The results on the standard partial molar volumes of transfer, compressibility and enthalpy of dilution from water to aqueous alcohol and diol solutions have been correlated and interpreted in terms of ion-polar, ion-hydrophobic, and hydrophobic-hydrophobic group interactions. The heat of dilution of these amino acids, peptides, and hen egg white lysozyme measured in aqueous solutions of 3-chloropropano-1-ol and 3-chloropropan-1,2-diol by using isothermal titration calorimetry along with the volumetric, compressibility