Glyoxysomal Malate Dehydrogenase in Pumpkin : Cloning of a cDNA and Functional Analysis of Its Presequence

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概要

• 論文の詳細を見る

• Glyoxysomal malate dehydrogenase (gMDH) is an enzyme of the glyoxylate cycle that participates in degradation of storage oil. We have cloned a cDNA for gMDH from etiolated pumpkin cotyledons that encodes a polypeptide consisting of 356 amino acid residues. The nucleotide and N-terminal amino acid sequences revealed that gMDH is synthesized as a precursor with an N-terminal extrapeptide. The N-terminal presequence of 36 amino acid residues contains two regions homologous to those of other microbody proteins, which are also synthesized as large precursors. To investigate the functions of the N-terminal presequence of gMDH, we generated transgenic Arabidopsis that expressed a chimeric protein consisting of β-blucuronidase and the N-terminal region of gMDH. Immunological and immunocytochemical studies revealed that the chimeric protein was imported into microbodies such as glyoxysomes and leaf peroxisomes and was then subsequently processed. Site-directed mutagensis studies showed that the conserved amino acids in the N-terminal presequence, Arg-10 and His-17, function as recognition sites for the targeting to plant microbodies, and Cys-36 in the presequence is responsible for its processing. These results correspond to those from the analyses of glyoxysomal citrate synthase (gCS), which was also synthesized as a large precursor, suggesting that common mechanisms that can recognize the targeting or the processing of gMDH and gCS function in higher plant cells.

• 日本植物生理学会の論文

著者

• NISHIMURA Mikio

  Department of Cell Biology, National Institute for Basic Biology

• HAYASHI Makoto

  Department of Basic Medical Research and Education, Ehime University Graduate School of Medicine

• KATO Akira

  Department of Biological Sciences, Tokyo Institute of Technology

• Moriya Kei

  Department Of Applied Biology Faculty Of Textile Science And Technology Shinshu University

• Hayashi M

  Department Of Cell Biology National Institute For Basic Biology

• Nishimura M

  Dep. Of Cell Biology National Inst. For Basic Biology

• Nishimura Mikio

  Department Of Cell Biology National Institute For Basic Biology

• Nishimura Mikio

  Dept. Cell Biol. Nat. Inst. Basic. Biol.

• Kato A

  Department Of Biology Faculty Of Science Niigata University

• KONDO Maki

  Department of Cell Biology, National Institute for Basic Biology

• Hara-Nishimura Ikuko

  Department of Botany, Graduate School of Science, Kyoto University

• Kojima M

  Department Of Bioresource Chemistry Obihiro University Of Agriculture And Veterinary Medicine

• HARA(NISHIMURA) Ikuko

  Department of Botany, Graduate School of Science, Kyoto University

• Hara-nishimura Ikuko

  Dept. Cell Biol. Nat. Inst. Basic. Biol.

• Hara Ikuko

  Department Of Botany Graduate School Of Science Kyoto University

• Hara-nishimura Ikuko

  Department Of Botany Graduate School Of Science Kyoto University

• Kojima Michiyuki

  Department Of Bioresource Chemistry Obihiro University Of Agriculture And Veterinary Medicine

• Kondo Maki

  Department Of Cell Biology National Institute For Basic Biology

• Takeda-yoshikawa Yoko

  Department Of Biology Faculty Of Science Kobe University

• Nishimura Mikio

  National Inst. For Basic Biology Okazaki Jpn

• Komai Michio

  Nof Corporation

• Kato Akira

  Department Of Biological Sciences Tokyo Institute Of Technology

• Hayashi Makoto

  Depart. Of Bioteclnology Graduate School Of Eng. Osaka Univ.

• Kato Akira

  Department Of Anesthesiology Kansai Medical University

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