Struktur-Funktionsbeziehungen von Katalase-Peroxidasen
Abstract
Peroxidasen katalysieren die vielfältigsten biochemischen Reaktionen. Neben der Entsorgung von toxischen Peroxiden in der Zelle sind sie an der Lignin Biosynthese (einem der häufigsten Biopolymere der Erde), an der unspezischen Immunabwehr des Menschen oder beispielsweise an Hormonbiosynthesen (z.B. in der Schilddrüse) beteiligt. Peroxidasen der Klasse I der Superfamilie I (dazu gehören Enzyme aus Bakterien, Pilzen und Pflanzen) sind äußerst interessante Studienobjekte der Enzymologie. Zu Klasse I Peroxidasen gehören bakterielle Katalase-Peroxidasen, Cytochrom c Peroxidase aus Hefe und Ascorbat Peroxidasen aus Pflanzen. Trotz sehr hoher Aminosäurehomologie zeigen diese 3 Vertreter hinsichtlich ihrer Reaktivität jedoch sehr starke Unterschiede. Die gemeinsame Funktion ist zwar die Entsorgung des intrazellulär gebildeten toxischen Wasserstoffperoxids, jedoch benötigt Cytochrom c Peroxidase dazu als Elektronendonor ein Protein (Cytochrom c), während Ascorbat Peroxidasen die Wasserstoffperoxid Reduktion mittels des hydrophilen Anions Ascorbinsäure durchführen. Und Katalase-Peroxidasen scheinen (und damit sind sie einzigartig unter Peroxidasen) Wasserstoffperoxid ähnlich den typischen Katalasen mit Hilfe eines weiteren Peroxid Moleküls unter Freisetzung von Wasser und Sauerstoff zu entsorgen. Daneben zeigen sie aber auch typische Peroxidase Aktivitäten und können hydrophobe aromatische Verbindungen oxidieren. Sie sind also bifunktionell (daher die Bezeichnung Katalase-Peroxidasen). Aufgrund dieser unterschiedlichen Reaktivität trotz hoher Homologie sind Klasse I Peroxidasen zur Untersuchung von Struktur-Funktionsbeziehungen optimal geeignet. Da der Mechanismus ihrer katalytischen Aktivität bislang unbekannt ist, werden in diesem Projekt rekombinante Proteine aus drei verschiedenen Bakterien untersucht. Durch ortsspezifische Mutagenese in Kombination mit verschiedenen spektroskopischen Methoden (UV-Vis Spektroskopie, Resonanz Raman Spektroskopie, Elektronen Spin Resonanz Spektroskopie) und kinetischen Untersuchungsmethoden (Multi-mixing Stopped-flow Spektroskopie, Polarographie) soll der Reaktionsmechanismus aufgeklärt werden und ein wertvoller Beitrag zum Verständnis von Hämproteinen (zu denen neben Peroxidasen auch Katalasen, Myoglobin, Hämoglobin, Cytochrome und viele andere im Stoffwechsel sehr wichtige Enzyme zählen) geleistet werden. Insbesondere ermöglichen diese Untersuchungsmethoden die Charakterisierung der Struktur und Reaktivität der Redoxintermediate dieser metallhältigen Enzyme und das Verständnis der Rolle der Aminosäuren der Proteinmatrix an der Substratbindung und -oxidation, am Elektronentransport zur prosthetischen Hämgruppierung und an der Wasserstoffperoxid Reduktion (und bei Katalase Aktivität auch Wasserstoffperoxid Oxidation). Da die dreidimensionale Struktur dieser Enzyme bislang unbekannt ist, sind zudem umfangreiche Kristallisationsversuche geplant.
Publications
Engineering the proximal heme cavity of catalase-peroxidase.
Autoren: Jakopitsch, C; Regelsberger, G; Furtmüller, PG; Rüker, F; Peschek, GA; Obinger, C Jahr: 2002
Journal articles
Understanding the Heme Cavity of Catalase-Peroxidase via Site-Directed Mutagenesis and Resonance Raman Spectroscopy
Autoren: Smulevich, G., Santoni, E., Heering, H. A., Indiani, C., Jakopitsch, C., Regelsberger, G., and Obinger, C. Jahr: 2002
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Distal site aspartate is essential in the catalase activity of catalase-peroxidases.
Autoren: Jakopitsch, C; Auer, M; Regelsberger, G; Jantschko, W; Furtmüller, PG; Rüker, F; Obinger, C Jahr: 2003
Journal articles
Identification of the radical intermediates formed by the catalase-peroxidase of Synechocystis PCC6803: A multifrequency EPR spectroscopy study combined with isotope labelling and site directed mutagenese
Autoren: Ivancich, A., Jakopitsch, C., and Obinger, C. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Towards understanding the catalatic activity of peroxidases and catalases?
Autoren: Jakopitsch, C., Schmuckenschlager, F., Zehner, F., Wanasinghe, A., Furtmüller, P. G., Obinger, C. Jahr: 2004
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:The role in the stability of the heme architecture of selected catalase-peroxidase mutants. An overview of a spectroscopic study.
Autoren: Droghetti, E., Jakopitsch, C., Obinger, C., Smulevich, G Jahr: 2005
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:The role of the unusual crosslinked distal site Trp-Tyr-Met adduct in catalysis of bifunctional catalase-peroxidase
Autoren: Jakopitsch, C., Vlasits, J., Ivancich, A., Obinger, C., Jahr: 2005
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Trapping the spectral features of the redox intermediates in the catalatic cycle of catalase-peroxidase
Autoren: Jakopitsch, C., Vlasits, J., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Identification of Trp106 as the tryptophanyl radical intermediate in Synechocystis PCC6803 catalase-peroxidase by multifrequency Electron Paramagnetic Resonance spectroscopy.
Autoren: Jakopitsch, C; Obinger, C; Un, S; Ivancich, A Jahr: 2006
Journal articles
Redox thermodynamics of the Fe(III)/Fe(II) couple in wild-type heme peroxidases and in their CN-adducts
Autoren: Bellei, M., Jakopitsch, C., Battistuzzi, G., Sola, M., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Protein-based radical intermediates in bifunctional heme peroxidases
Autoren: Ivancich, A., Un, S., Obinger, C., Loewen, P. Jahr: 2005
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Mechanism of reaction of horseradish peroxidase with chlorite and chlorine dioxide.
Autoren: Jakopitsch, C; Spalteholz, H; Furtmüller, PG; Arnhold, J; Obinger, C; Jahr: 2008
Journal articles
Modulation of catalytic functions in heme enzymes by posttranslational modifications of redox sites
Autoren: Obinger, C. Jahr: 2011
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Chlorite as oxidant and reductant of heme peroxidases: a mechanistic study
Autoren: Furtmüller, P.G., Jakopitsch, C., Spalteholz, H., Arnhold, J., Obinger, C. Jahr: 2007
Journal articles
external links and characteristics of the publication:Do we really understand the catalatic reactivity?
Autoren: Jakopitsch, C; Petutschnig, G; Zehner, F; Ruker, F; Furtmuller, PG; Obinger, C Jahr: 2003
Journal articles
Interaction with the Redox Cofactor MYW and Functional Role of a Mobile Arginine in Eukaryotic Catalase-Peroxidase
Autoren: Gasselhuber, B; Graf, MMH; Jakopitsch, C; Zamocky, M; Nicolussi, A; Furtmuller, PG; Oostenbrink, C; Carpena, X; Obinger, C Jahr: 2016
Journal articles
New insights into the heme cavity structure of catalase-peroxidase: a spectroscopic approach to the recombinant synechocystis enzyme and selected distal cavity mutants.
Autoren: Heering, HA; Indiani, C; Regelsberger, G; Jakopitsch, C; Obinger, C; Smulevich, G Jahr: 2002
Journal articles
The 2.0A resolution structure of the catalytic portion of a cyanobacterial membrane-bound manganese superoxide dismutase.
Autoren: Atzenhofer, W; Regelsberger, G; Jacob, U; Peschek, G; Furtmüller, P; Huber, R; Obinger, C Jahr: 2002
Journal articles
The catalytic role of the distal site asparagine-histidine couple in catalase-peroxidases.
Autoren: Jakopitsch, C; Auer, M; Regelsberger, G; Jantschko, W; Furtmüller, PG; Rüker, F; Obinger, C Jahr: 2003
Journal articles
Spectroscopic and kinetic investigations of selected mutants of recombinant catalase-peroxidase from synecocystis
Autoren: Santoni, E., Jakopitsch, C., Heering, H.A., Indiani, C., Regelsberger, G., Obinger, C., Smulevich, G., Jahr: 2003
Chapter in collected volumes
external links and characteristics of the publication:Total conversion of bifunctional catalase-peroxidase (KatG) to monofunctional peroxidase by exchange of a conserved distal side tyrosine.
Autoren: Jakopitsch, C; Auer, M; Ivancich, A; Rüker, F; Furtmüller, PG; Obinger, C Jahr: 2003
Journal articles
Distal side tryptophan, tyrosine and methionine in catalase-peroxidases are covalently linked in solution.
Autoren: Jakopitsch, C; Kolarich, D; Petutschnig, G; Furtmüller, PG; Obinger, C Jahr: 2003
Journal articles
Comparison between catalase-peroxidase and cytochrom c peroxidase: A resonance raman study
Autoren: Santoni, E., Jakopitsch, C., Obinger, C., and Smulevich, G. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Engineering catalase-peroxidases: The structural basis of the activity of catalase-peroxidases.
Autoren: Petutschnig, G., Jakopitsch, C., Auer, M., Rüker, F., Furtmüller, P. G., and Obinger, C. Jahr: 2003
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Heme peroxidase biochemistry--facts and perspectives.
Autoren: Obinger, C Jahr: 2010
Journal articles
Mechanism of hydrogen peroxide oxidation in catalase-peroxidases
Autoren: Jakopitsch, C., Vlasits, J., Obinger, C. Jahr: 2010
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Protein-based radicals in the catalase-peroxidase of synechocystis PCC6803: a multifrequency EPR investigation of wild-type and variants on the environment of the heme active site.
Autoren: Ivancich, A; Jakopitsch, C; Auer, M; Un, S; Obinger, C Jahr: 2003
Journal articles
Insights into catalase-peroxidase by electronic absorption, resonance Raman spectroscopy, and site-directed mutagenesis
Autoren: Smulevich, G., Santoni, E., Jakopitsch, C., Obinger, C. Jahr: 2004
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Comparison between catalase-peroxidase and cytochrome c peroxidase. The role of the hydrogen-bond networks for protein stability and catalysis.
Autoren: Santoni, E; Jakopitsch, C; Obinger, C; Smulevich, G Jahr: 2004
Journal articles
Spectral and kinetic studies on redox intermediates in heme peroxidases from both superfamilies
Autoren: Obinger, C., Furtmüller, P.G., Zederbauer, M., Jakopitsch, C. Jahr: 2005
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:The role of selected residues in the heme architecture of catalase-peroxidases
Autoren: Droghetti, E., Jakopitsch, C., Obinger, C., Smulevich, G. Jahr: 2005
Conference & Workshop proceedings, paper, abstract
Probing the role of the peculiar Trp-Tyr-Met adduct in the heme pocket of bifunctional catalase-peroxidases
Autoren: Jakopitsch, C., Vlasits, J., Ivancich, A., Obinger, C. Jahr: 2005
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Probing the structure and bifunctionality of catalase-peroxidase (KatG).
Autoren: Smulevich, G; Jakopitsch, C; Droghetti, E; Obinger, C Jahr: 2006
Journal articles
Redox thermodynamics of the ferric-ferrous couple of wild-type synechocystis KatG and KatG(Y249F).
Autoren: Bellei, M; Jakopitsch, C; Battistuzzi, G; Sola, M; Obinger, C Jahr: 2006
Journal articles
The catalatic activity of monofunctional catalases and bifunctional catalase-peroxidases follows different mechanisms
Autoren: Obinger, C., Vlasits, J., Jakopitsch, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Investigation of the catalytic mechanism of Synechocystis catalase-peroxidase with doubly O18-labeled hydrogen peroxide
Autoren: Vlasits, J., Jakopitsch, C., Holubar, P., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Identification of the unique site (Trp106) for the formation of the [Fe(IV)=O Trp.] intermediate in Synechocystis PCC6803 KatG and the (distal heme side) extended H-bonding network with a crucial role in catalase activity vy using multifrequency (9-285 GHz) EPR spectroscopy
Autoren: Jakopitsch, C., Obinger, C., Un, S., Ivancich, A. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Towards understanding the role of protein-based radical intermediates as alternative oxidation sites in mono- and bifunctional peroxidases (KatGs)
Autoren: Loewen, P. C., Obinger, C., Un, S., Ivancich, A. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Detoxification of hydrogen peroxide in cyanobacteria
Autoren: Bernroitner, M., Jakopitsch, C., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Probing the reaction of three distinct catalase-peroxidases with peroxides
Autoren: Jakopitsch, C., Wiseman, B., Vlasits, J., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Hydrogen peroxide oxidation by catalase-peroxidase follows a non-scrambling mechanism.
Autoren: Vlasits, J; Jakopitsch, C; Schwanninger, M; Holubar, P; Obinger, C; Jahr: 2007
Journal articles
Catalases
Autoren: Obinger, C., Furtmüller, P. G., Zamocky, M. Jahr: 2012
Chapter in collected volumes
Investigation of the catalytic mechanism of Synechocystis catalase-peroxidase with doubly O18-labeled hydrogen peroxide
Autoren: Vlasits, J., Jakopitsch, C., Holubar, P., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:A GC-MS study on the catalatic mechanism of catalase-peroxidase
Autoren: Vlasits, J., Jakopitsch, C., Holubar, P., Obinger, C. Jahr: 2006
Conference & Workshop proceedings, paper, abstract
external links and characteristics of the publication:Mechanisms of catalase activity of heme peroxidases.
Autoren: Vlasits, J; Jakopitsch, C; Bernroitner, M; Zamocky, M; Furtmüller, PG; Obinger, C; Jahr: 2010
Journal articles
Redox thermodynamic of the Fe3+/Fe2+ couple in wild-type and mutated heme peroxidases
Autoren: Battistuzzi, G., Bellei, M., Jakopitsch, C., Vlasits, J., Furtmüller, P.G., Sola, M., Obinger, C. Jahr: 2007
Journal articles
external links and characteristics of the publication:Fungal catalase-peroxidases - a novel group of bifunctional oxidoreductases
Autoren: Zamocky, M., Jakopitsch, C., Vlasits, J., Obinger, C. Jahr: 2007
Journal articles
external links and characteristics of the publication:
Project staff
Christian Obinger
Univ.Prof. Mag.rer.nat. Dr.rer.nat. Christian Obinger
christian.obinger@boku.ac.at
Tel: +43 1 47654-10011, 77273
Projektleiter*in
01.01.2002 - 31.12.2004
Christa Jakopitsch
Mag.Dr. Christa Jakopitsch
Tel: +43 1 47654-33061
Projektmitarbeiter*in
01.01.2002 - 31.12.2004
Martina Paumann-Page
Dipl.-Ing. Dr.rer.nat. Martina Paumann-Page
Projektmitarbeiter*in
01.01.2002 - 31.12.2004
BOKU partners
External partners
Institute of Chemistry
Giulietta Smulevich
Partner
Section Bioenergetics, Biophysics of Oxidative Stress_x000D_ CEA-Saclay, France
Anabella Ivancich
Partner