| 1. |
- Henriksson, Gunnar, et al.
(författare)
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Is cellobiose dehydrogenase from Phanerochaete chrysosporium a lignin degrading enzyme?
- 2000
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Ingår i: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. - 0167-4838 .- 1879-2588. ; 1480:02-jan, s. 83-91
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Tidskriftsartikel (refereegranskat)abstract
- Cellobiose dehydrogenase (CDH) is an extracellular redox enzyme of ping-pong type, i.e. it has separate oxidative and reductive half reactions. Several wood degrading fungi produce CDH, but the biological function of the enzyme is not known with certainty. It can, however, indirectly generate hydroxyl radicals by reducing Fe3+ to Fe2+ and O-2 to H2O2. Hydroxyl radicals are then generated by a Fenton type reaction and they can react with various wood compounds, including lignin. In this work we study the effect of CDH on a non-phenolic lignin model compound (3,4-dimethoxyphenyl glycol). The results indicate that CDH can affect lignins in three important ways. (1) It breaks beta-ethers; (2) it demethoxylates aromatic structures in lignins; (3) it introduces hydroxyl groups in non-phenolic lignins. The gamma-irradiated model compound gave a similar pattern of products as the CDH treated model compound? when the samples were analyzed by HPLC, suggesting that hydroxyl radicals are the active component of the CDH system.
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| 2. |
- Ottosson, Jenny, et al.
(författare)
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Size as a parameter for solvent effects on Candida antarctica lipase B enantioselectivity
- 2002
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Ingår i: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. - 0167-4838 .- 1879-2588. ; 1594:2, s. 325-334
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Tidskriftsartikel (refereegranskat)abstract
- Changes in solvent type were shown to yield significant improvement of enzyme enantioselectivity. The resolution of 3-methyl-2-butanol catalyzed by Candida antarctica lipase B, CALB, was studied in eight liquid organic solvents and supercritical carbon dioxide, SCCO2. Studies of the temperature dependence of the enantiomeric ratio allowed determination of the enthalpic (Delta(R-S)Delta H-double dagger) as well as the entropic (Delta(R-S)Delta S-double dagger) contribution to the overall enantioselectivity (Delta(R-S)Delta G(double dagger) = -RTlnE). A correlation of the enantiomeric ratio, E. to the van der Waals volume of the solvent molecules was observed and suggested as one of the parameters that govern solvent effects on enzyme catalysis. An enthalpy-entropy compensation relationship was indicated between the studied liquid solvents. The enzymatic mechanism must be of a somewhat different nature in SCCO2, as this reaction in this medium did not follow the enthalpy-entropy compensation relation.
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| 3. |
- Kleczkowski, Leszek A, 1954-
(författare)
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Is leaf ADP-glucose pyrophosphorylase an allosteric enzyme?
- 2000
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Ingår i: Biochimica et Biophysica Acta - Protein Structure and Molecular Enzymology. - 0167-4838 .- 1879-2588. ; 1476:1, s. 103-108
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Tidskriftsartikel (refereegranskat)abstract
- Barley leaf ADP-glucose pyrophosphorylase (AGPase), a key enzyme of starch synthesis in the chloroplast stroma, was analysed, in both directions of the reaction, with respect to details of its regulation by 3-phosphoglycerate (PGA) and inorganic phosphate (Pi) which serve as activator and inhibitor, respectively. AGPase was found to catalyse a close-to-equilibrium reaction, with the K-eq value of approximately 0.5, i.e. slightly favouring the pyrophosphorolytic direction. When the enzyme was analysed by substrate kinetics, PGA acted either as a linear (hyperbolic response) 'non-competitive' activator (forward reaction) or a linear near-'competitive' activator (reverse reaction). When the activation and inhibition patterns with PGA and Pi, respectively, were studied in detail by Dixon plots, the response curves to effecters also followed hyperbolic kinetics, with the experimentally determined K-a and K-i values on the order of micromolar. The results suggest that the regulation of AGPase proceeds via a non-cooperative mechanism, where neither of the effecters, when considered separately, induces any allosteric response. The evidence, discussed in terms of an overall kinetic mechanism/regulation of leaf AGPase, prompts caution in classifying the protein as an 'allosteric enzyme'. (C) 2000 Elsevier Science B.V. All rights reserved.
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| 4. |
- Berggren, Kristina, et al.
(författare)
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Substitutions of surface amino acid residues of cutinase probed by aqueous two-phase partitioning
- 2000
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Ingår i: BBA - Protein Structure and Molecular Enzymology. - 0167-4838. ; 1481:2, s. 317-327
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Tidskriftsartikel (refereegranskat)abstract
- The surface properties of a protein are often crucial for recognition and interaction with other molecules. Important functional residues can be identified by mutational analysis. There is a need for rapid methods to study protein surfaces and surface changes due to mutations. Partitioning in aqueous two-phase systems has the potential to be used in this respect since protein partitioning depends on the surface properties of the protein. The influence of surface-exposed amino acid residues in protein partitioning has been studied with cutinase variants, which differed in one or several amino acid residues as a result of site-directed mutagenesis. The solvent accessibility of the mutated residues was determined with a computer program, Graphical Representation and Analysis of Surface Properties. The aqueous two-phase system was composed of dextran and a random copolymer of ethylene oxide and propylene oxide. It was shown, for the first time, to what extent surface-exposed amino acid residues influence the partition coefficient in an aqueous two-phase system. The effect on partitioning could be described only taking into account solvent accessibility and type of residue substitution. The results demonstrate that the system can be used to detect conformational changes in mutant proteins since the expected effect on partitioning due to a mutation can be calculated. The aqueous two-phase system used here does indeed provide a rapid and convenient method to study protein surfaces and slight surface changes due to mutations.
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| 5. |
- Berggren, K, et al.
(författare)
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The surface exposed amino acid residues of monomeric proteins determine the partitioning in aqueous two-phase systems
- 2002
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Ingår i: BBA - Protein Structure and Molecular Enzymology. - 0167-4838. ; 1596:2, s. 253-268
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Tidskriftsartikel (refereegranskat)abstract
- It is of great interest and importance how different amino acid residues contribute to and affect the properties of a protein surface, Partitioning in aqueous two-phase systems has the potential to be used as a rapid and simple method for studying the surface properties of proteins. The influence on partitioning of the surface exposed amino acid residues of eight structurally determined monomeric proteins has been studied. The proteins were characterized in terms of surface exposed residues with a computer program, Graphical Representation and Analysis of Surface Properties (GRASP), and partitioned in two EO30PO70-dextran aqueous two-phase systems, only differing in polymer concentrations (system I: 6.8% EO30PO70, 7.1% dextran; system II: 9% EO30PO70, 9% dextran). We show for the first time that the partitioning behaviour of different monomeric proteins can be described by the differences in surface exposed amino acid residues. The contribution to the partition coefficient of the residues was found to be best characterized by peptide partitioning in the aqueous two-phase system. Compared to hydrophobicity scales available in the literature, each amino acid contribution is characterized by the slope given by the graph of log K against peptide chain length, for peptides of different length containing only one kind of residue. It was also shown that each amino acid contribution is relative to the total protein surface and the other residues on the surface. Surface hydrophobicity calculations realized for systems I and 11 gave respectively correlation coefficients of 0.961 and 0.949 for the linear relation between log K and calculated hydrophobicity values. To study the effect on the partition coefficient of different amino acids, they were grouped into classes according to common characteristics: the presence of an aromatic group, a long aliphatic chain or the presence of charge. Using these groups it was possible to confirm that aromatic residues have the strongest effect on the partition coefficient, giving preference to the upper EO30PO70 phase of the system; on the other hand the presence of charged amino acids on the protein surface enhances the partition of the protein to the lower dextran phase. It is also important to note that the sensitivity of the EO30PO70-dextran system for the surface exposed residues was increased by increasing the polymer concentrations. The partition coefficient of a monomeric protein can thus be predicted from its surface exposed amino acid residues and the system can also be used to characterize protein surfaces of monomeric proteins in general. (C) 2002 Elsevier Science B.V. All rights reserved.
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| 6. |
- Härndahl, Ulrika, et al.
(författare)
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The chaperone-like activity of a small heat shock protein is lost after sulfoxidation of conserved methionines in a surface-exposed amphipathic α-helix
- 2001
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Ingår i: BBA - Protein Structure and Molecular Enzymology. - 0167-4838. ; 1545:1-2, s. 227-237
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Tidskriftsartikel (refereegranskat)abstract
- The small heat shock proteins (sHsps) possess a chaperone-like activity which prevents aggregation of other proteins during transient heat or oxidative stress. The sHsps bind, onto their surface, molten globule forms of other proteins, thereby keeping them in a refolding competent state. In Hsp21, a chloroplast-located sHsp in all higher plants, there is a highly conserved region forming an amphipathic α-helix with several methionines on the hydrophobic side according to secondary structure prediction. This paper describes how sulfoxidation of the methionines in this amphipathic α-helix caused conformational changes and a reduction in the Hsp21 oligomer size, and a complete loss of the chaperone-like activity. Concomitantly, there was a loss of an outer-surface located α-helix as determined by limited proteolysis and circular dichroism spectroscopy. The present data indicate that the methionine-rich amphipathic α-helix, a motif of unknown physiological significance which evolved during the land plant evolution, is crucial for binding of substrate proteins and has rendered the chaperone-like activity of Hsp21 very dependent on the chloroplast redox state.
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| 7. |
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| 8. |
- Henriksson, G, et al.
(författare)
-
Is cellobiose dehydrogenase from Phanerochaete chrysosporium a lignin degrading enzyme?
- 2000
-
Ingår i: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY. - 0167-4838. ; 1480:1-2, s. 83-91
-
Tidskriftsartikel (refereegranskat)abstract
- Cellobiose dehydrogenase (CDH) is an extracellular redox enzyme of ping-pong type, i.e. it has separate oxidative and reductive half reactions. Several wood degrading fungi produce CDH, but the biological function of the enzyme is not known with certainty
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| 9. |
- Ridderstrom, M, et al.
(författare)
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The active-site residue Tyr-175 in human glyoxalase II contributes to binding of glutathione derivatives
- 2000
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Ingår i: BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY. - 0167-4838. ; 1481:2, s. 344-348
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Tidskriftsartikel (refereegranskat)abstract
- Tyrosine-175 located in the active site of human glyoxalase II was replaced by phenylalanine in order to study the contribution of this residue to catalysis. The mutation had a marginal effect on the k(cat) value determined using S-D-lactoylglutathione as
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| 10. |
- Biedron, Sandra G., et al.
(författare)
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The sensitivity of nonlinear harmonic generation to electron beam quality in free electron lasers
- 2002
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Ingår i: Nuclear Instruments & Methods in Physics Research. Section A: Accelerators, Spectrometers, Detectors, and Associated Equipment. - 0167-5087. ; 483:1-2, s. 101-106
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Tidskriftsartikel (refereegranskat)abstract
- The generation of harmonics through a nonlinear mechanism driven by bunching at the fundamental has sparked interest as a path toward enhancing and extending the usefulness of an X-ray free-electron laser (FEL) facility. The sensitivity of the nonlinear harmonic generation to undulator imperfections, electron beam energy spread, peak current, and emittance is important in an evaluation of the process. Typically, linear instabilities in FELs are characterized by increased sensitivity to both electron beam and undulator quality with increasing harmonic number. However, since the nonlinear harmonic generation mechanism is driven by the growth of the fundamental, the sensitivity of the nonlinear harmonic mechanism is not expected to be significantly greater than that of the fundamental. In this paper, we study the effects of electron beam quality, more specifically, emittance, energy spread, and peak current, on the nonlinear harmonics in a 1.5-Angstrom FEL, and show that the decline in the harmonic emission roughly follows that of the fundamental. (C) 2002 Elsevier Science B.V. All rights reserved.
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