Structure/Function Studies on Flavoproteins

Summary

Principal Investigator: Irina Sevrioukova
Abstract: [unreadable] DESCRIPTION (provided by applicant): The aim of the proposed work is to use a combination of structural, biochemical and genetic approaches to study structure/function relationships in flavoproteins. The work will focus on two structurally homologous FAD-containing enzymes, putidaredoxin reductase from Pseudomonas putida and apoptosis inducing factor from mice. Putidaredoxin reductase catalyzes electron transfer from NADH to an iron-sulfur protein, putidaredoxin, in cytochrome P450cam monooxygenase. The mechanism of complex formation and electron transfer between putidaredoxin reductase and putidaredoxin is not well understood. We have recently demonstrated that putidaredoxin reductase has redox active cysteines and can function as dithiol/disulfide oxidoreductase. Since there are no putative disulfide redox centers encoded by CysXXCys or CysXXXXCys motifs characteristic for traditional disulfide reductases, the mechanism of dithiol/disulfide oxidoreduction catalyzed by putidaredoxin reductase seems to be unique and needs to be elucidated. Apoptosis inducing factor is a phylogenetically old mammalian, caspase-independent death effector which, upon apoptosis induction, translocates from its normal localization, the mitochondrial intermembrane space, to the nucleus where it causes chromatin condensation and DNA fragmentation. Neither physiological function nor mechanism of apoptosis induced by this protein is known. [unreadable] [unreadable] Our research will address the question of how the structures of putidaredoxin reductase and [unreadable] apoptosis inducing factor are related to their catalytic function. Crystal structures of putidaredoxin reductase and apoptosis inducing factor will be solved, compared, and related to the catalytic properties of the enzymes. From structural interpretations, these relationships will be further probed and modified by genetic engineering and the effect of specific structural changes on catalytic function of the proteins will be assessed. The study will explain the mechanisms of electron transfer and dithiol/disulfide oxidoreduction catalyzed by putidaredoxin reductase and will provide an insight into the mechanism and function of apoptosis inducing factor. [unreadable] [unreadable]
Funding Period: 2003-08-01 - 2009-07-31
more information: NIH RePORT

Top Publications

  1. pmc Cowchock syndrome is associated with a mutation in apoptosis-inducing factor
    Carlo Rinaldi
    Neurogenetics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892 3705, USA
    Am J Hum Genet 91:1095-102. 2012
  2. pmc Redox reactions of the FAD-containing apoptosis-inducing factor (AIF) with quinoidal xenobiotics: a mechanistic study
    Lina Miseviciene
    Institute of Biochemistry of Vilnius University, Mokslininku, Lithuania
    Arch Biochem Biophys 512:183-9. 2011
  3. pmc Structural biology of redox partner interactions in P450cam monooxygenase: a fresh look at an old system
    Irina F Sevrioukova
    Departments of Molecular Biology and Biochemistry, Chemistry and Pharmaceutical Sciences, University of California, Irvine, CA 92697 3900, USA
    Arch Biochem Biophys 507:66-74. 2011
  4. ncbi Arginines 65 and 310 in putidaredoxin reductase are critical for interaction with putidaredoxin
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    Biochemistry 49:5160-6. 2010
  5. pmc Crystal structure of the putidaredoxin reductase x putidaredoxin electron transfer complex
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    J Biol Chem 285:13616-20. 2010
  6. ncbi Production and characterization of a functional putidaredoxin reductase-putidaredoxin covalent complex
    Inna Y Churbanova
    Department of Molecular Biology, University of California, Irvine, California 92697 3900, USA
    Biochemistry 49:58-67. 2010
  7. pmc Redox-linked conformational dynamics in apoptosis-inducing factor
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, 92697 3900, USA
    J Mol Biol 390:924-38. 2009
  8. ncbi Redox-dependent changes in molecular properties of mitochondrial apoptosis-inducing factor
    Inna Y Churbanova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    J Biol Chem 283:5622-31. 2008
  9. pmc Putidaredoxin-to-cytochrome P450cam electron transfer: differences between the two reductive steps required for catalysis
    Vadim Yu Kuznetsov
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    Biochemistry 45:11934-44. 2006
  10. ncbi The putidaredoxin reductase-putidaredoxin electron transfer complex: theoretical and experimental studies
    Vadim Yu Kuznetsov
    Department of Molecular Biology, University of California, Irvine, California 92612 3900, USA
    J Biol Chem 280:16135-42. 2005

Scientific Experts

  • Irina Sevrioukova
  • Inna Y Churbanova
  • Vadim Yu Kuznetsov
  • Thomas L Poulos
  • Carlo Rinaldi
  • Lina Miseviciene
  • Massimo Zeviani
  • Costanza Lamperti
  • Kenneth H Fischbeck
  • Marina L Kennerson
  • Leslie G Biesecker
  • Iren Horkayne-Szakaly
  • Daniele Ghezzi
  • Barrington G Burnett
  • Christopher Grunseich
  • Alice Schindler
  • Guida Landoure
  • Carsten Bonnemann
  • Jonas Sarlauskas
  • Narimantas Cenas
  • Zilvinas Anusevicius
  • Amanda Pifferitti
  • Emek Blair
  • Patrick J Farmer

Detail Information

Publications10

  1. pmc Cowchock syndrome is associated with a mutation in apoptosis-inducing factor
    Carlo Rinaldi
    Neurogenetics Branch, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892 3705, USA
    Am J Hum Genet 91:1095-102. 2012
    ..Our findings expand the spectrum of AIF-related disease and provide insight into the effects of AIFM1 mutations...
  2. pmc Redox reactions of the FAD-containing apoptosis-inducing factor (AIF) with quinoidal xenobiotics: a mechanistic study
    Lina Miseviciene
    Institute of Biochemistry of Vilnius University, Mokslininku, Lithuania
    Arch Biochem Biophys 512:183-9. 2011
    ..Thus, these compounds may prevent the accumulation of the reduced form of AIF in vivo, and enhance AIF-mediated apoptosis...
  3. pmc Structural biology of redox partner interactions in P450cam monooxygenase: a fresh look at an old system
    Irina F Sevrioukova
    Departments of Molecular Biology and Biochemistry, Chemistry and Pharmaceutical Sciences, University of California, Irvine, CA 92697 3900, USA
    Arch Biochem Biophys 507:66-74. 2011
    ..The insights gained from these studies will be summarized in this review...
  4. ncbi Arginines 65 and 310 in putidaredoxin reductase are critical for interaction with putidaredoxin
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    Biochemistry 49:5160-6. 2010
    ....
  5. pmc Crystal structure of the putidaredoxin reductase x putidaredoxin electron transfer complex
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    J Biol Chem 285:13616-20. 2010
    ....
  6. ncbi Production and characterization of a functional putidaredoxin reductase-putidaredoxin covalent complex
    Inna Y Churbanova
    Department of Molecular Biology, University of California, Irvine, California 92697 3900, USA
    Biochemistry 49:58-67. 2010
    ....
  7. pmc Redox-linked conformational dynamics in apoptosis-inducing factor
    Irina F Sevrioukova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, 92697 3900, USA
    J Mol Biol 390:924-38. 2009
    ..Structural findings agree with biochemical data and support the hypothesis that both normal and apoptogenic functions of AIF are controlled by NADH...
  8. ncbi Redox-dependent changes in molecular properties of mitochondrial apoptosis-inducing factor
    Inna Y Churbanova
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    J Biol Chem 283:5622-31. 2008
    ....
  9. pmc Putidaredoxin-to-cytochrome P450cam electron transfer: differences between the two reductive steps required for catalysis
    Vadim Yu Kuznetsov
    Department of Molecular Biology and Biochemistry, University of California, Irvine, California 92697 3900, USA
    Biochemistry 45:11934-44. 2006
    ....
  10. ncbi The putidaredoxin reductase-putidaredoxin electron transfer complex: theoretical and experimental studies
    Vadim Yu Kuznetsov
    Department of Molecular Biology, University of California, Irvine, California 92612 3900, USA
    J Biol Chem 280:16135-42. 2005
    ....