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Structural biology of carboxylases in C4 plant

Research Project

Project/Area Number 13450353
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 工業物理化学
Research InstitutionOsaka University

Principal Investigator

KAI Yasushi  Grad. Sch. Engineering, Dept. Materials Chemistry, Professor, 大学院・工学研究科, 教授 (40029236)

Co-Investigator(Kenkyū-buntansha) MATSUMURA Hiroyoshi  Grad. Sch. Engineering, Dept. Materials Chemistry, Assistant Professor, 大学院・工学研究科, 助手 (30324809)
KANEHISA Nobuko  Grad. Sch. Engineering, Dept. Materials Chemistry, Associate Professor, 大学院・工学研究科, 講師 (20177538)
INOUE Tsuvoshi  Grad. Sch. Engineering, Dept. Materials Chemistry, Associate Professor, 大学院・工学研究科, 助教授 (20263204)
MOCHIZUKI Eiko  Grad. Sch. Engineering, Dept. Materials Chemistry, Research Assistant, 大学院・工学研究科, 教務員 (10150335)
Project Period (FY) 2001 – 2002
Project Status Completed (Fiscal Year 2002)
Budget Amount *help
¥15,000,000 (Direct Cost: ¥15,000,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2001: ¥12,800,000 (Direct Cost: ¥12,800,000)
KeywordsC4 plant / Maize / Carboxylase / PEP carboxylase / Effector / PEPC-Protein Kinase / X-ray analysis / Molecular mechanism
Research Abstract

Phosphoenolpyruvate carboxylase (PEPC) catalyzes the first step in the fixation of atmospheric CO_2 during C_4 photosynthesis. The crystal structure of C_4-form maize PEPC, the first structure of the plant PEPCs, has been determined at 3.0 A^^○ resolution. The structure includes a sulfate ion at the plausible binding site of an allosteric activator, glucose 6-phosphate. The crystal structure of E. coli PEPC complexed with Mn2^<2+>, phosphoenolpyruvate analogue (3,3-dichloro-2-dihydroxyphosphinoylmethyl-2-propenoate), and an allosteric inhibitor, aspartate, has also been determined at 2.35 A^^○ resolution. Dynamic movements were found in the maize PEPC structure around two loops near the active site, compared with E. coli PEPC. Based on these molecular structures, the mechanisms for the carboxylation reaction and for the allosteric regulation of PEPC are proposed.
Ribulose 1,5-bisphosphate carboxylase/oxygenase (RuBisCO) is the initial step enzyme in the Calvin-Benson cycle of photosynthesis. It catalyzes the addition of gaseous CO_2 to ribulose 1,5-bisphosphate and produces two molecules of 3-phosphoglycerate. However, this enzyme also catalyzes O_2 addition for RuBP as the primary reaction of photorespiration. The crystal structure of Rubisco from Chlamydomonas has also been determined at 1.84 A^^○ resolution. The present high-resolution structure has revealed novel post-translational modifications. The crystal structure of unactivated Galdieria Rubisco has also been determined at 2.6 A^^○ resolution. The electron density map reveals that a sulfate binds only to the P1 anion-binding site of the active site and the loop 6 is closed. This interaction is likely to be crucial to understanding for stabilizing the loop 6 in the close state and to making a higher affinity for anionic ligands.

Report

(3 results)
  • 2002 Annual Research Report   Final Research Report Summary
  • 2001 Annual Research Report
  • Research Products

    (21 results)

All Other

All Publications (21 results)

  • [Publications] H.Matsumura, et al.: "Crystal structures of active state C_4-form maize and quaternary complex of E.coli phosphoenolpyruvate carboxylases : Implications for the mechanisms of the carboxylation reaction and allosteric regulation"Structure. 10. 1721-1730 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Inoue, et al.: "Metal Activation Mechanism of Human Hematopoietic Prostaglandin D Synthase"Nat.Struct.Biol.. 10. 291-296 (2003)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T.Yamamoto, et al.: "Crystallization and preliminary X-ray analysis of TBP-interacting protein from the hyperthermophilic archaeon Thermococcus kodakaraensis strain KOD1"Acta Crystallogr.. D59. 372-374 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Kai, et al.: "Phosphoenolpyruvate carboxylase : Three-dimensional structure and molecular mechanisms"Arch.Biochem.Biophys.. (in press).

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y.Okano, et al.: "X-ray structure of Galdieria Rubisco complexed with one sulfate ion per active site"FEBS Lett.. 527. 33-36 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] E.Mizohata, et al.: "Crystal structure of activated ribulose-1,5-biphosphate carboxylase/oxygenase from green alga Chlamydomonas reinhardtii complexed with 2-carboxyarabinitol-1,5-biphosphate"J.Mol.Biol.. 316. 679-691 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H. Matsumura, et al.: "Crystal structures of active state C_4-form maize and quaternary complex of E. coli phosphoenolpyruvate carboxylases : Implications for the mechanism a of the carboxylation reaction and allosteric regulation"Structure. 10. 1721-1730 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Inoue, et al.: "Metal Activation Mechanism of Human Hematopoietic Prostaglandin D Syntheae"Nat. Siruct. Blol.. 10. 291-296 (2003)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] T. Yamamoto, et al.: "Crystallization and preliminary X-ray analysis of TBP-interacting protein from the hyperthermophilic archaeon Thermococcus kodakaraensis strain KO DI"Acta Crystallogr.. D59. 372-374 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y. Kai, et al.: "Phosphoenolpyruvate carboxylase : Three-dimensional structure and moledular mechanisms"Arch. Biochem. Biophys.. in press.

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] Y. Okano, et al.: "X-ray structure of Galdieria Rubisco complexed with one sulfate ion per active site"FEBS Lett.. 527. 33-36 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] E. Mizohata, et al.: "Crystal structure of activated ribulose-1,5-biphosphate carboxylase/oxygenase from green alga Chlamydomonas reinhardtii complexed with 2-carboxyara binitol-1,5-biphosphate"J. Mol. Biol.. 316. 679-691 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2002 Final Research Report Summary
  • [Publications] H.Matsumura, et al.: "Crystal structures of active state C_4-form maize and quaternary complex of E. coli phosphoenolpyruvate carboxylases : Implications for the mechanisms of the carboxylation reaction and allosteric regulation"Structure. 10. 1721-1730 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Inoue, et al.: "Metal Activation Mechanism of Human Hematopoietic Prostaglandin D Synthase"Nat.Struct.Biol.. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] T.Yamamoto, et al.: "Crystallization and preliminary X-ray analysis of TBP-interacting protein from the hyperthermophilic archaeon Thermococcus kodakaraensis strain KOD1"Acta Crystallogr.. D59. 372-374 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Y, Kai, et al.: "Phosphoenolpyruvate carboxylase : Three-dimensional structure and molecular mechanisms"Arch.Biochem.Biophys.. (in press).

    • Related Report
      2002 Annual Research Report
  • [Publications] Y.Okano, et al.: "X-ray structure of Galdieria Rubisco complexed with one sulfate ion per active site"FEBS Lett.. 527. 33-36 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] E.Mizohata, et al.: "Crystal structure of activated ribulose-1,5-biphoshate carboxylase/oxygenase from green alga Chlamydomonas reinhardtii complexed with 2-carboxyarabinitol-1,5-biphosphate"J.Mol.Biol.. 316. 679-691 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] E.Mizohata, et al.: "Crystallization and preliminary X-ray diffraction analysis of glutathione-dependent dehydroascorbate reductase from spinach chloroplasts"Acta Cryst.. D57. 1726-1728 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] H.Hashimoto, et al.: "Crystal structure of DNA polymerase from hyperthermophilic archaeon Pyrococcus Kodakaraensis KOD1"J. Mol. Biol.. 306. 469-477 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] E.Mizohata, et al.: "Crystal structure of activated ribulose-1, 5-bisphosphate carboxylase/oxygenase from green alga Chlamydomonas reinhardtii complexed with 2-cafboxyarabinitol-1, 5-bisphosphate"J. Mol. Biol.. 316. 677-689 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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