Isolation and proteomic analysis of rice Golgi membranes : cis-Golgi membranes labeled with GFP-SYP31

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概要

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• The Golgi membranes labeled with cis-Golgi marker GFP-SYP31 were isolated from suspension-cultured cells of rice transformed with 35S::GFP-SYP31 by floating through a discontinuous sucrose density gradient in the presence of 5 mM MgCl2. The specific fluorescence intensity of final membrane preparation increased to approximately 150-fold in comparison with that of the post-nucleus soluble fraction. Specific activity of membrane-bound α-mannosidase (cis-Golgi) markedly increased, but NDPase (medial-/trans-Golgi) and NADPH-cytochrome c reductase endoplasmic reticulum were weakly detected in the membrane fraction. The other organelle marker enzymes, cytochrome c oxidase (mitochondria), alkaline pyrophosphatase (plastid), and catalase (peroxisome) were not detectable. Comparative display of protein spots in GFP-SYP31-labeled membranes, microsomal membranes and soluble fraction on the two dimensional gels showed that some proteins are markedably concentrated in the cis-Golgi membrane fraction. Furthermore, the mass spectrometric analysis of proteins separated by SDS-polyacrylamide gel electrophoresis indicated that the highly purified Golgi membranes contained several membrane traffic-related proteins and ER resident proteins. These findings indicated a close relationship between the ER and the cis-Golgi membranes.

• 2006-12-01

著者

• Hori Hidetaka

  Graduate School of Science and Technology, Niigata University

• Mitsui Toshiaki

  Graduate School of Science and Technology, Niigata University

• Hori H

  Graduate School Of Sci. And Technol. Niigata Univ.

• Hori Hidetaka

  Laboratories Of Molecular Life Science Graduate School Of Science And Technology Niigata University

• Hori Hidetaka

  Laboratories Of Molecular Life Sciences Graduate School Of Natural Science And Technology Niigata Un

• Hori Hidetaka

  Graduate School Of Natural Science And Technology Niigata University

• Mitsui Toshiaki

  Laboratories of Molecular Life Sciences, Graduate School of Science and Technology, Niigata Universi

• SHIMAMOTO Ko

  Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology

• Sato Masa

  Faculty of Integrated Human Studies, Kyoto University

• Mitsui T

  Graduate School Of Science And Technology Niigata University

• Mitsui Toshiaki

  Laboratories Of Molecular Life Science Graduate School Of Science And Technology Niigata University

• Mitsui Toshiaki

  Graduate School Of Natural Science And Technology Niigata University

• FUJIWARA Masayuki

  Laboratory of Plant Molecular Genetics, Nara Institute of Science and Technology

• ASAKURA Tsuyoshi

  Laboratories of Plant and Mircobial Genome Control, Graduate School of Science and Technology, Niiga

• HIROSE Shota

  Laboratories of Plant and Mircobial Genome Control, Graduate School of Science and Technology, Niiga

• KATAMINE Hiroki

  Laboratories of Plant and Mircobial Genome Control, Graduate School of Science and Technology, Niiga

• KITAJIMA Aya

  Laboratories of Plant and Mircobial Genome Control, Graduate School of Science and Technology, Niiga

• KITAJIMA Aya

  Department of Applied Biological Chemistry, Niigata University

• Shimamoto Ko

  Laboratory For Plant Molecular Genetics Nara Institute Of Science And Technology

• ASAKURA Tsuyoshi

  Graduate School of Science and Technology, Niigata University

• HIROSE Shota

  Graduate School of Science and Technology, Niigata University

• Kitajima Aya

  Laboratories Of Plant And Mircobial Genome Control Graduate School Of Science And Technology Niigata

• Hirose Shota

  Graduate School Of Science And Technology Niigata University

• Fujiwara Masayuki

  Laboratory Of Plant Molecular And Plant Protein Analysis Plant Science Education Unit Graduate Schoo

• Sato Masa

  Faculty Of Human Environmental Sciences Kyoto Prefectural University

• Mitsui Toshiaki

  Laboratories Of Plant And Mircobial Genome Control Graduate School Of Science And Technology Niigata

• Hori Hidetaka

  Laboratories Of Plant And Mircobial Genome Control Graduate School Of Science And Technology Niigata

• Asakura Tsuyoshi

  Graduate School Of Science And Technology Niigata University

• Katamine Hiroki

  Laboratories Of Plant And Mircobial Genome Control Graduate School Of Science And Technology Niigata University

• Shimamoto Ko

  Nara Inst. Of Sci. And Technol. Ikoma Jpn

• Fujiwara Masayuki

  Laboratory of Neuropharmacology, Faculty of Pharmaceutical Sciences, Hiroshima International University, Japan

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