Distal site aspartate is essential in the catalase activity of catalase-peroxidases.


:Structural and biochemical characterization of aspartate 152 at the distal heme side of catalase-peroxidase (KatG) from Synechocystis PCC 6803 reveals an important functional role for this residue. In the wild-type protein, the side chain carboxyl group of Asp152 is 7.8 A apart from the heme iron and is hydrogen-bonded to two water molecules and a KatG-specific large loop. We have prepared the site-specific variants Asp152Asn, Asp152Ser, Asp152Trp, and Pro151Ala. Exchange of Asp152 exhibited dramatic consequences on the bifunctional activity of this unique peroxidase. The turnover number of catalase activity of Asp152Asn is 2.7%, Asp152Ser 5.7%, and Asp152Trp is 0.6% of wild-type activity. By contrast, the peroxidase activity of the Asp152 variants was 2-7 times higher than that of wild-type KatG or Pro151Ala. The KatG-specific pH profile of the catalase activity was completely different in these variants and exchange of Asp152 made it possible to follow the transition of the ferric enzyme to the redox intermediate compound I by hydrogen peroxide spectroscopically and to determine the corresponding bimolecular rate constant to be 7.5 x 10(6) M(-1) s(-1) (pH 7 and 15 degrees C). The reactivity of compound I toward aromatic one-electron donors was enhanced in the Asp152 variants compared with the wild-type protein, whereas the reactivity toward hydrogen peroxide was dramatically decreased. A mechanism for the hydrogen peroxide oxidation, which is different from monofunctional catalases and involves the distal residues Trp122 and Asp152, is proposed.






Jakopitsch C,Auer M,Regelsberger G,Jantschko W,Furtmüller PG,Rüker F,Obinger C




Has Abstract


2003-05-13 00:00:00












  • Identification of essential tyrosine and lysine residues in angiotensin converting enzyme: evidence for a single active site.

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    pub_type: 杂志文章


    authors: Chen YN,Riordan JF

    更新日期:1990-11-20 00:00:00

  • Main chain and side chain dynamics of a heme protein: 15N and 2H NMR relaxation studies of R. capsulatus ferrocytochrome c2.

    abstract::A detailed characterization of the main chain and side chain dynamics in R. capsulatus ferrocytochrome c(2) derived from (2)H NMR relaxation of methyl group resonances is presented. (15)N relaxation measurements confirm earlier results indicating that R. capsulatus ferrocytochrome c(2) exhibits minor rotational anisot...


    pub_type: 杂志文章


    authors: Flynn PF,Bieber Urbauer RJ,Zhang H,Lee AL,Wand AJ

    更新日期:2001-06-05 00:00:00

  • The EcoRI-DNA complex as a model for investigating protein-DNA interactions by atomic force microscopy.

    abstract::Atomic force microscopy (AFM) is a technique widely used to image protein-DNA complexes, and its application has now been extended to the measurements of protein-DNA binding constants and specificities. However, the spreading of the protein-DNA complexes on a flat substrate, generally mica, is required prior to AFM im...


    pub_type: 杂志文章


    authors: Sorel I,Piétrement O,Hamon L,Baconnais S,Cam EL,Pastré D

    更新日期:2006-12-12 00:00:00

  • Crystal structure of the acid-induced arginine decarboxylase from Escherichia coli: reversible decamer assembly controls enzyme activity.

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    pub_type: 杂志文章


    authors: Andréll J,Hicks MG,Palmer T,Carpenter EP,Iwata S,Maher MJ

    更新日期:2009-05-12 00:00:00

  • Factors affecting gamma-chain multimer formation in cross-linked fibrin.

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    pub_type: 杂志文章


    authors: Siebenlist KR,Mosesson MW

    更新日期:1992-01-28 00:00:00

  • New protein cross-linking reagents that are cleaved by mild acid.

    abstract::New homo- and heterobifunctional cross-linking reagents have been synthesized. These reagents are based on ortho ester, acetal, and ketal functionalities that undergo acid-catalyzed dissociation but are base stable. The protein-reactive group in all the homobifunctional reagents is a maleimide group; the heterobifunct...


    pub_type: 杂志文章


    authors: Srinivasachar K,Neville DM Jr

    更新日期:1989-03-21 00:00:00

  • Generation and Comparative Kinetic Analysis of New Glycosynthase Mutants from Streptococcus pyogenes Endoglycosidases for Antibody Glycoengineering.

    abstract::Chemoenzymatic glycan remodeling by endoglycosidase-catalyzed deglycosylation and reglycosylation is emerging as an attractive approach for producing homogeneous glycoforms of antibodies, and the success of this approach depends on the discovery of efficient endoglycosidases and their glycosynthase mutants. We report ...


    pub_type: 杂志文章


    authors: Tong X,Li T,Li C,Wang LX

    更新日期:2018-09-04 00:00:00

  • Phosphorylation of protein phosphatase 2Czeta by c-Jun NH2-terminal kinase at Ser92 attenuates its phosphatase activity.

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    pub_type: 杂志文章


    authors: Awano K,Amano K,Nagaura Y,Kanno S,Echigo S,Tamura S,Kobayashi T

    更新日期:2008-07-08 00:00:00

  • Measurement of DNA-protein equilibria using gel chromatography: application to the HinfI restriction endonuclease.

    abstract::A method is described for measuring equilibrium constants of DNA-protein interactions using gel chromatography. This technique has been used to study the sequence-specific interaction of the HinfI restriction endonuclease with DNA. HinfI has a monomeric molecular weight of 31000 and exists as a dimer in its active for...


    pub_type: 杂志文章


    authors: Frankel AD,Ackers GK,Smith HO

    更新日期:1985-06-04 00:00:00

  • The relationship between ligand aggregation and G-quadruplex DNA selectivity in a series of 3,4,9,10-perylenetetracarboxylic acid diimides.

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    pub_type: 杂志文章


    authors: Kern JT,Thomas PW,Kerwin SM

    更新日期:2002-09-24 00:00:00

  • Mobilities of the inner three core residues and the Man(alpha 1--6) branch of the glycan at Asn78 of the alpha-subunit of human chorionic gonadotropin are restricted by the protein.

    abstract::Human chorionic gonadotropin (hCG) is a heterodimeric glycoprotein hormone involved in the maintenance of the corpus luteum in early pregnancy. The free alpha-subunit of hCG has a biological activity of its own, namely, stimulation of prolactin secretion from term pregnancy decidual cells [Blithe, D. L., et al. (1991)...


    pub_type: 杂志文章


    authors: van Zuylen CW,de Beer T,Leeflang BR,Boelens R,Kaptein R,Kamerling JP,Vliegenthart JF

    更新日期:1998-02-17 00:00:00

  • Structural Snapshots of an Engineered Cystathionine-γ-lyase Reveal the Critical Role of Electrostatic Interactions in the Active Site.

    abstract::Enzyme therapeutics that can degrade l-methionine (l-Met) are of great interest as numerous malignancies are exquisitely sensitive to l-Met depletion. To exhaust the pool of methionine in human serum, we previously engineered an l-Met-degrading enzyme based on the human cystathionine-γ-lyase scaffold (hCGL-NLV) to cir...


    pub_type: 杂志文章


    authors: Yan W,Stone E,Zhang YJ

    更新日期:2017-02-14 00:00:00

  • Stopped-flow analysis on anion binding to blue-form halorhodopsin from Natronobacterium pharaonis: comparison with the anion-uptake process during the photocycle.

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    pub_type: 杂志文章


    authors: Sato M,Kanamori T,Kamo N,Demura M,Nitta K

    更新日期:2002-02-19 00:00:00

  • Pressure effects on dipalmitoylphosphatidylcholine bilayers measured by 2H nuclear magnetic resonance.

    abstract::The effects of pressure, up to 5 kbar, on multilamellar vesicles of 1,2-dipalmitoyl-sn-phosphatidylcholine perdeuterated in the acyl chains (DPPC-d62) were examined by using high-pressure NMR techniques. A deuterium probe was built, and the quadrupole splitting was measured against pressure at various temperatures. Th...


    pub_type: 杂志文章


    authors: Driscoll DA,Samarasinghe S,Adamy S,Jonas J,Jonas A

    更新日期:1991-04-02 00:00:00

  • Temperature dependence of forward and reverse electron transfer from A1-, the reduced secondary electron acceptor in photosystem I.

    abstract::Electron-transfer reactions following the formation of P700(+)A1- have been studied in isolated Photosystem I complexes from Synechococcus elongatus between 300 and 5 K by flash absorption spectroscopy. (1) In the range from 300 to 200 K, A1- is reoxidized by electron transfer to the iron-sulfur cluster FX. The rate s...


    pub_type: 杂志文章


    authors: Schlodder E,Falkenberg K,Gergeleit M,Brettel K

    更新日期:1998-06-30 00:00:00

  • Specificity determinants on Cdc42 for binding its effector protein ACK.

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    pub_type: 杂志文章


    authors: Elliot-Smith AE,Mott HR,Lowe PN,Laue ED,Owen D

    更新日期:2005-09-20 00:00:00

  • The 0.78 A structure of a serine protease: Bacillus lentus subtilisin.

    abstract::Ultrahigh-resolution X-ray diffraction data from cryo-cooled, B. lentus subtilisin crystals has been collected to a resolution of 0.78 A. The refined model coordinates have a rms deviation of 0.22 A relative to the same structure determined at room temperature and 2.0 A resolution. Several regions of main-chain and si...


    pub_type: 杂志文章


    authors: Kuhn P,Knapp M,Soltis SM,Ganshaw G,Thoene M,Bott R

    更新日期:1998-09-29 00:00:00

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    pub_type: 杂志文章


    authors: Halling SM,Sanchez-Anzaldo FJ,Fukuda R,Doi RH,Meares CF

    更新日期:1977-06-28 00:00:00

  • Cold adaptation of enzyme reaction rates.

    abstract::A major issue for organisms living at extreme temperatures is to preserve both stability and activity of their enzymes. Cold-adapted enzymes generally have a reduced thermal stability, to counteract freezing, and show a lower enthalpy and a more negative entropy of activation compared to mesophilic and thermophilic ho...


    pub_type: 杂志文章


    authors: Bjelic S,Brandsdal BO,Aqvist J

    更新日期:2008-09-23 00:00:00

  • A kinetic study of interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates with type II bovine cardiac muscle adenosine cyclic 3',5'-phosphate dependent protein kinase.

    abstract::The stereoselectivity of the adenosine cyclic 3',5'-phosphate (cAMP) binding sites on the regulatory subunit of the type II bovine cardiac muscle cAMP-dependent protein kinase was investigated by examining the interactions of (Rp)- and (Sp)-adenosine cyclic 3',5'-phosphorothioates (cAMPS) with these sites. While activ...


    pub_type: 杂志文章


    authors: O'Brian CA,Roczniak SO,Bramson HN,Baraniak J,Stec WJ,Kaiser ET

    更新日期:1982-08-31 00:00:00

  • Apical membrane of the gastric parietal cell: water, proton, and nonelectrolyte permeabilities.

    abstract::Gastric parietal cell apical membranes must protect the cell from the extremely low pH and wide variations in osmolality of the gastric juice. To characterize the permeability properties of gastric apical membranes, we have measured passive permeabilities to water, protons, NH3, and small nonelectrolytes of membrane v...


    pub_type: 杂志文章


    authors: Priver NA,Rabon EC,Zeidel ML

    更新日期:1993-03-16 00:00:00

  • A conformational rearrangement in gramicidin A: from a double-stranded left-handed to a single-stranded right-handed helix.

    abstract::A conformational transition is described for the polypeptide, gramicidin A, in which a dimer that forms a left-handed intertwined antiparallel helix is converted to a single-stranded amino terminus to amino terminus right-handed helix. The starting structure is determined here by solution NMR methods while reference i...


    pub_type: 杂志文章


    authors: Zhang Z,Pascal SM,Cross TA

    更新日期:1992-09-22 00:00:00

  • Ligand Design for Specific MHC Class I Molecules on the Cell Surface.

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    pub_type: 杂志文章


    authors: Sun X,Tokunaga R,Nagai Y,Miyahara R,Kishimura A,Kawakami S,Katayama Y,Mori T

    更新日期:2020-12-15 00:00:00

  • Determination of singlet oxygen-specific versus radical-mediated lipid peroxidation in photosensitized oxidation of lipid bilayers: effect of beta-carotene and alpha-tocopherol.

    abstract::Photosensitized oxidation reactions damage tissue by catalyzing the formation of oxyradicals and singlet oxygen. beta-Carotene is hypothesized to exert photoprotective effects by quenching singlet oxygen formed by Type II reactions and by scavenging free radicals formed by Type I reactions. beta-Carotene antioxidant m...


    pub_type: 杂志文章


    authors: Stratton SP,Liebler DC

    更新日期:1997-10-21 00:00:00

  • Insight into the mechanism of serpin-proteinase inhibition from 2D [1H-15N] NMR studies of the 69 kDa alpha 1-proteinase inhibitor Pittsburgh-trypsin covalent complex.

    abstract::We have used [(1)H-(15)N]-HSQC NMR to investigate the structural changes that occur in both serpin and proteinase in forming the kinetically trapped covalent protein-protein complex that is the basis for serpin inhibition of serine proteinases. By alternately using (15)N-alanine specifically-labeled alpha(1)-proteinas...


    pub_type: 杂志文章


    authors: Peterson FC,Gettins PG

    更新日期:2001-05-29 00:00:00

  • SecA insertion into phospholipids is stimulated by negatively charged lipids and inhibited by ATP: a monolayer study.

    abstract::SecA-lipid interactions are believed to be important for the translocation of precursor proteins across the inner membrane of Escherichia coli [Lill, R., Dowhan, W., & Wickner, W. (1990) Cell 60, 271-280]. SecA insertion into the phospholipid bilayer could a role in this process. We investigated this possibility by st...


    pub_type: 杂志文章


    authors: Breukink E,Demel RA,de Korte-Kool G,de Kruijff B

    更新日期:1992-02-04 00:00:00

  • Crystal structure of eta-crystallin: adaptation of a class 1 aldehyde dehydrogenase for a new role in the eye lens.

    abstract::Eta-crystallin is a retinal dehydrogenase that has acquired a role as a structural protein in the eye lens of elephant shrews, members of an ancient order of mammals. While it retains some activity toward retinal, which is oxidized to retinoic acid, the protein has acquired a number of specific sequence changes that h...


    pub_type: 杂志文章


    authors: Bateman OA,Purkiss AG,van Montfort R,Slingsby C,Graham C,Wistow G

    更新日期:2003-04-22 00:00:00

  • Backbone Flexibility Influences Nucleotide Incorporation by Human Translesion DNA Polymerase η opposite Intrastrand Cross-Linked DNA.

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    pub_type: 杂志文章


    authors: O'Flaherty DK,Guengerich FP,Egli M,Wilds CJ

    更新日期:2015-12-29 00:00:00

  • A fluorescent pentameric thiophene derivative detects in vitro-formed prefibrillar protein aggregates.

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    pub_type: 杂志文章


    authors: Hammarström P,Simon R,Nyström S,Konradsson P,Aslund A,Nilsson KP

    更新日期:2010-08-17 00:00:00

  • Distinct activation states of alpha5beta1 integrin show differential binding to RGD and synergy domains of fibronectin.

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    pub_type: 杂志文章


    authors: García AJ,Schwarzbauer JE,Boettiger D

    更新日期:2002-07-23 00:00:00