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2001 Fiscal Year Final Research Report Summary

Structure/function analysis of abundance and transmission mechanism of genetic information invlolved in the biosynthesis of multi-catalytic site enzyme

Research Project

Project/Area Number 11460039
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

MURATA Kousaku  Kyoto University, Agriculture, Professor, 農学研究科, 教授 (90142299)

Co-Investigator(Kenkyū-buntansha) KAWAI Shigeyuki  Kyoto University, Agriculture, Research Associate, 農学研究科, 助手 (00303909)
HASHIMOTO Wataru  Kyoto University, Agriculture, Associate Professor, 農学研究科, 助教授 (30273519)
Project Period (FY) 1999 – 2001
Keywordssphingomonas / alginate lyase / autoprocessing / protease / x-ray crystallography / β-elimination mechanism
Research Abstract

The intracellular alginate lyases I,II, and III are coded by one gene and produced through sequential post-translational reactions from a common pre-cursor protein Po : Po → I + 5 kDa peptide, I → II +III. The protease-like activity of I is completely inhibited by EDTA or O- phnonthroline, thus indicates that the I is a multi- catalytic site enzyme currying II, III, and protease. The protease site on I is thought to be created through the excision of N-terminal 5 kDa peptide from Po, since Po has no protease activity. An important biological problem is then raised as to how the protease site is generated and how / where the information is described in the base and amino acid sequences. The clarification of these addresses may lead to the postulation of a novel theory concerning the abundance of information contained in a gene. To solve the problems, the knowleges on the structure / function relationship of alginate lyases I, II, and III, as well as their precursor protein Po. Among these, the structure / function of III was completely analyzed through X-ray crystallography and detailed lyase reaction mechanism was clarified. Especially, a novel β-elimination reaction mechanism involving Tyr residue, but not His, was constructed with an emphasis of Low-Barrier Hydrogen Bond Theory.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Yumiko Mishima: "Super-channel in bacteria : macromolecule uptake and depolymerization systems of Sphingomonas sp. A1 with special cell surface structure"Biotechnology & Genetic Engineering Reviews. (in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Keiko Momma: "Crystal structure of AlgQ2, a macromolecule(alginate)-binding protein of Sphingomonas sp. A1 at 2.0Å resoltion"J. Mol. Biol.. 316(5). 1061-1069 (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hye-Jin Yoon: "Crystal structure of alginate lyase A1-III complexed with trisaccharide product at 2.0Å resolution"J. Mol. Biol.. 307. 9-16 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hye-Jin Yoon: "Effect of His192 mutation on the activity of alginate lyase A1-III from Sphingomanas species A1"J. Microbiol. biotechnol.. 11(1). 9-16 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yumiko Mishima: "Crystallization and preliminary X-ray analysis of ΛlgS, a bacterial ATP-binding cassette(ABC) protein specific to macromole-cule import"Acta Cryst.. D57. 884-885 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wataru Hashimoto: "Polysaccharide lyase : molecular cloning, sequencing, and overexpression of the xanthan lyase gene of Bacillus sp. strain GL1"Appl. Environ. Microbiol.. 67(2). 713-720 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Wataru Hashimoto: "Super-channel in bacteria : Function and structure of a macromolecule import system mediated by a pit-dependent ABC transporter"Biosci. Biotechnol. Biochem.. 65(9). 1949-1956 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yumiko Mishima: "Super-channel in bacteria : Function and structure of the macromolecule import system mediated by a pit-dependent ABC transporter"FEMS Microbiol. Lett.. 204. 215-221 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yumiko Mishima: "Super-channel in bacteria : macromolecule uptake and depolymerization systems of Sphingomonas sp. A1 with special cell surface structure"Biotechnology & Genetic Egineering Reviews. (in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Keiko Momma: "Crystal structure of AlgQ2, a macromolecule (alginate)-binding protein of Sphingomonas sp. A1 at 2.0A resolution"J. Mol. Biol.. 316(5). 1061-1069 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hye-Jin Yoon: "Crystal structure of alginate lyase A1-III complexed with trisaccharide product at 2.0A resolution"J. Mol. Biol.. 307. 9-16 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Hye-Jin Yoon: "Effect of His192 mutation on the activity of alginate lyase A1-III from Sphingomonas spedes A1"J. Microbiol. Biotechnol.. 11(1). 118-123 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yumiko Mishima: "Crystallization and preliminary X-ray analysis of AlgS, a bacterial ATP-binding cassette(ABC) protein specific to macromolecule import"Acta Cryst.. D57. 884-885 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wataru Hashimoto: "Polysaccharide lyase : molecular cloning, sequencing, and overexpression of the xanthan lyase gene of Bacillus sp. Strain GL1"Appl. Environ. Microbiol.. 67(2). 713-720 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Wataru Hashimoto: "Super-channel in bacteria : Function and structure of a macromolecule import system medeated by a pit-dependent ABC transporter"Biosci. Biotechnol. Biochem.. 65(9). 1949-1956 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yumiko Mishima: "Super-channel in bacteria : Function and structure of the macromolicule import system mediated by a pit-dependent ABC transporter"FEMS Microbiol. Lett.. 204. 215-221 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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