徳島大学 教育・研究者情報データベース(EDB)

1. 徳島大学.ゲノム機能研究センター(〜2008年3月31日/->組織[徳島大学.疾患プロテオゲノム研究センター])

1. 山本 武範

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2. 山田 安希子

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3. (英) Watanabe Masahiro

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4. (英) Yoshimura Yuya

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5. 山﨑 尚志([徳島大学.大学院医歯薬学研究部.薬学域.薬科学部門.創薬科学系.衛生薬学]/[徳島大学.薬学部.創製薬科学科.創薬学講座.衛生薬学]/[徳島大学.薬学研究科.創薬科学専攻])

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6. 吉村 好之

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7. 山内 卓

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8. 片岡 正俊

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9. 永田 俊彦

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10. 寺田 弘([東京理科大学])

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11. 篠原 康雄([徳島大学.先端酵素学研究所.基幹研究部門])

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(英) VDAC1, having a shorter N-terminus than VDAC2 but showing the same migration in an SDS-polyacrylamide gel, is the predominant form expressed in mitochondria of various tissues.

(英) The voltage-dependent anion channel (VDAC) is a pore-forming protein expressed in the outer membrane of eukaryotic mitochondria. Three isoforms of it, i.e., VDAC1, VDAC2, and VDAC3, are known to be expressed in mammals; however, the question as to which is the main isoform in mitochondria is still unanswered. To address this question, we first prepared standard VDACs by using a bacterial expression system and raised various antibodies against them by using synthetic peptides as immunogens. Of the three bacterially expressed VDAC isoforms, VDAC3 showed faster migration in SDS-polyacrylamide gels than VDAC1 and VDAC2, although VDAC2 is longer than VDAC1 and VDAC3, due to a 12-amino acid extension of its N-terminal region. Even with careful structural characterization of the expressed VDACs by LC-MS/MS analysis, serious structural modifications of VDACs causing changes in their migration in SDS-polyacrylamide gels were not detected. Next, immunoreactivities of the raised antibodies toward these bacterially expressed VDAC isoforms were evaluated. Trials to prepare specific antibodies against the three individual VDAC isoforms were not successful except in the case of VDAC1. However, using a synthetic peptide corresponding to the highly conserved region among the three VDACs, we were successful in preparing an antibody showing essentially equal immunoreactivities toward all three VDACs. When mitochondrial outer membrane proteins of various rat tissues were subjected to 2-dimensional electrophoresis followed by immunoblotting with this antibody, six immunoreactive protein spots were detected. These spots were characterized by LC-MS/MS analysis, and the signal intensities among the spots were compared. As a result, the signal intensity of the spot representing VDAC1 was the highest, and thus, VDAC1 was concluded to be the most abundantly expressed of the three VDAC isoforms in mammalian mitochondria.

1. (英) Amino Acid Sequence
2. (英) Animals
3. (英) Base Sequence
4. (英) Chromatography, Liquid
5. (英) DNA Primers
6. (英) Electrophoresis, Gel, Two-Dimensional
7. (英) Electrophoresis, Polyacrylamide Gel
8. (英) Immunoblotting
9. (英) Mass Spectrometry
10. (英) Mitochondrial Proteins
11. (英) Molecular Sequence Data
12. (英) Protein Isoforms
13. (英) Rats
14. (英) Sequence Analysis, DNA
15. (英) Voltage-Dependent Anion Channel 1

1. (英) Article.Affiliation: Institute for Genome Research, University of Tokushima, Kuramotocho-3, Tokushima 770-8503, Japan.

2. (英) Article.PublicationTypeList.PublicationType: Comparative Study

3. (英) Article.PublicationTypeList.PublicationType: Journal Article