Characterization of arsenic-binding proteins and its application to health effect evaluation.

• Project/Area Number: 23390167
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Hygiene
• Research Institution: National Institute for Environmental Studies
• Principal Investigator: HIRANO Seishiro 独立行政法人国立環境研究所, 環境リスク研究センター, 室長 (20150162)
• Co-Investigator(Kenkyū-buntansha): KOBAYASHI Yayoi 独立行政法人国立環境研究所, 環境健康研究センター, 主任研究員 (00391102)
• Project Period (FY): 2011-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥17,550,000 (Direct Cost: ¥13,500,000、Indirect Cost: ¥4,050,000); Fiscal Year 2013: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2012: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2011: ¥7,540,000 (Direct Cost: ¥5,800,000、Indirect Cost: ¥1,740,000)
• Keywords: 環境中毒 / ヒ素 / タンパク質 / 電気泳動 / promyelocytic leukemia / SUMO化 / 蛍光染色 / キャラクタリゼーション / 遺伝子導入 / 細胞 / 毒性 / Promyelocytic leukemia / 結合タンパク質 / 担体

Research Abstract

We prepared arsenic-bound sepharoses and investigated arsenic-binding proteins by chromatography using a column packed with the arsenic-bound sepharoses. SDS-PAGE analyses showed no proteins bound to pentavalent arsenic specifically, while several proteins that have specific and high affinity for trivalent arsenic. We prepared promyelocytic leukemia (PML) expressing cells and applied the cytosolic proteins to the arsenic-binding column and the column was elutes with glutathione-containing solution. PML appeared to bind trivalent arsenic by forming ternary complex at RING finger domain of this protein. Western blot and immunofluorescent microscopic analyses suggested that PML were conjugation with small ubiquitin-like modifier proteins (SUMOylation) in response to arsenic exposure. The arsenic-bound sepharose column enabled us to identify proteins which have a high affinity to arsenic using this method.

Research Products

• [Journal Article] Metabolism of Arsenic and Its Toxicological Relevance (2013)
  • Author(s): TakayukiWatanabe and Seishiro Hirano
  • Journal Title: Arch. Toxicol Volume: 87 Pages: 969-979
  • Peer Reviewed
• [Journal Article] The role of glutathione in the metabolism of diphenylarsinic acid in rats (2013)
  • Author(s): Kobayashi, Y. and Hirano, S
  • Journal Title: Matallomics Volume: 5 Pages: 469-478
  • Peer Reviewed
• [Journal Article] MitochondrialElectron Transport is Inhibited by Disappearance of Metallothionein in Human Bronchial Epithelial Cells Following Exposure to Silver Nitrate (2013)
  • Author(s): Takamitsu Miyayama, Yuta Arai, Noriyuki Suzuki and Seishiro Hirano
  • Journal Title: Toxicology Volume: 305 Pages: 20-29
  • Peer Reviewed
• [Journal Article] Effects of arsenic on modification of promyelocytic leukemia (PML) : PML responds to low levels of arsenite (2013)
  • Author(s): Seishiro Hirano, TakayukiWatanabe, and Yayoi Kobayashi
  • Journal Title: Toxicol.Appl.Pharmacol Volume: 273(3) Issue: 3 Pages: 590-599
  • DOI: 10.1016/j.taap.2013.10.004
  • Peer Reviewed
• [Journal Article] Developmental subchronic exposure to diphenylarsinic acid induced increased exploratory behavior, impaired learning behavior, and decreased cerebellar glutathione concentration in rats (2013)
  • Author(s): Negishi T, Matsunaga Y, Kobayashi Y, Hirano S, Tashiro T
  • Journal Title: Toxicol. Sci Volume: 136(2) Pages: 478-86
  • Peer Reviewed
• [Journal Article] Metabolism of Arsenic and Its Toxicological Relevance. (2013)
  • Author(s): Takayuki Watanabe
  • Journal Title: Arch. Toxicol. Volume: 87 Issue: 6 Pages: 969-979
  • DOI: 10.1007/s00204-012-0904-5
  • Peer Reviewed
• [Journal Article] Mitochondrial Electron Transport is Inhibited by Disappearance of Metallothionein in Human Bronchial Epithelial Cells Following Exposure to Silver Nitrate. (2013)
  • Author(s): Takamitsu Miyayama
  • Journal Title: Toxicology Volume: 305 Pages: 20-29
  • DOI: 10.1016/j.tox.2013.01.004
  • Peer Reviewed
• [Journal Article] Diphenylarsinic Acid Increased Synthesis and Release of Neuroactive and Vasoactive Peptides in Rat Cerebellar Astrocytes (2012)
  • Author(s): Negishi T, Takahashi M, Matsunaga Y, Hirano S, Tomoko T
  • Journal Title: Journal of Neuropathology and Experimental Neurology Volume: 71 Pages: 468-479
  • Peer Reviewed
• [Journal Article] Diphenylarsinic Acid Increased Synthesis and Release of Neuroactive and Vasoactive Peptides in Rat Cerebellar Astrocytes.. (2012)
  • Author(s): Negishi T
  • Journal Title: Journal of Neuropathology and Experimental Neurology Volume: 71 Issue: 6 Pages: 468-479
  • DOI: 10.1097/nen.0b013e3182561327
  • Peer Reviewed
• [Journal Article] Analysis of arsenic metabolites in HepG2 and AS3MT-transfected cells (2011)
  • Author(s): TakayukiWatanabe, Yuki Ohta, Ayano Mizumura, Yayoi Kobayashi, and Seishiro Hirano
  • Journal Title: Arch. Toxicol Volume: 85 Pages: 577-588
  • Peer Reviewed
• [Journal Article] Rapid speciation and quantification of selenium compounds by HPLC-ICPMS using multiple standards labelled with different isotopes (2011)
  • Author(s): Yuki Ohta, Noriyuki Suzuki, Yayoi Kobayashi, and Seishiro Hirano
  • Journal Title: Isotopes in Environmental & Health Studies Volume: 43 Pages: 330-340
  • Peer Reviewed
• [Journal Article] Analysis of arsenic metabolites in HepG2 and AS3MT-transfected cells (2011)
  • Author(s): Takayuki Watanabe, Yuki Ohta, Ayano Mizumura, Yayoi Kobayashi, Seishiro Hirano
  • Journal Title: Arch.Toxicol. Volume: 85 Issue: 6 Pages: 577-588
  • DOI: 10.1007/s00204-011-0710-5
  • Peer Reviewed
• [Presentation] Tissue distribution and excretion of arsenic and selenium following co-administration of diphenylarsinic acid and selenite in rats (2014)
  • Author(s): Kobayashi Y., Hirano S
  • Organizer: 53rd SOTAnnual Meeting
  • Place of Presentation: Phoenix
• [Presentation] 2013)亜ヒ酸を曝露した細胞におけるPMLタンパク質の核内局在化 (2013)
  • Author(s): 平野靖史郎, 小林弥生, 渡辺喬之
  • Organizer: 第83回日本衛生学会
  • Place of Presentation: 金沢
• [Presentation] 多剤耐性関連タンパク質2 依存型・非依存型ヒ素の胆汁排泄に及ぼすセレンの影響 (2013)
  • Author(s): 小林弥生, 平野靖史郎
  • Organizer: フォーラム2013衛生薬学・環境トキシコロジー
  • Place of Presentation: 福岡
• [Presentation] 亜ヒ酸にPMLのSUMO化はAREの活性化を介さない (2013)
  • Author(s): 平野靖史郎, 小林弥生, 渡辺喬之, 伹野美保子
  • Organizer: 第19回ヒ素シンポジウム
  • Place of Presentation: 福岡
• [Presentation] Implication of relationship between arsenic toxicity and arsenic metabolites (2013)
  • Author(s): Yayoi Kobayashi, Ayano Mizumura, Ayaka Yamashiro, and Seishiro Hirano
  • Organizer: X ISTERH 2013 Journal of Trace Elements in Medicine and Biology
  • Place of Presentation: Tokyo
• [Presentation] 亜ヒ酸にPMLのSUMO化はAREの活性化を介さない. (2013)
  • Author(s): 平野靖史郎,小林弥生,渡辺喬之、伹野美保子
  • Organizer: 第19回ヒ素シンポジウム
  • Place of Presentation: 福岡
• [Presentation] 亜ヒ酸を曝露した細胞におけるPMLタンパク質の核内局在化 (2013)
  • Author(s): 平野靖史郎
  • Organizer: 第83回日本衛生学会
  • Place of Presentation: 金沢
• [Presentation] 多剤耐性タンパク質MRPsを介した細胞のヒ素毒性調節機構 (2013)
  • Author(s): 渡辺喬之
  • Organizer: 第83回日本衛生学会
  • Place of Presentation: 金沢
• [Presentation] 多剤耐性タンパク質MRP2高発現細胞における細胞内ヒ素代謝・排泄機構の解析 (2012)
  • Author(s): 渡辺喬之、小林弥生、平野靖史郎
  • Organizer: 第82回日本衛生学会総会
  • Place of Presentation: 京都大学(京都府)
  • Year and Date: 2012-03-25
• [Presentation] 多剤耐性タンパク質MRP2 高発現細胞における細胞内ヒ素代謝・排泄機構の解析 (2012)
  • Author(s): 渡辺喬之, 小林弥生, 平野靖史郎
  • Organizer: 第82回日本衛生学会総会
  • Place of Presentation: 京都
• [Presentation] Effects of arsenite on macropinocytotic and autophagic activities in CHO-K1 cells (2012)
  • Author(s): S. Hirano, T. Watanabe, Y. Kobayashi, S. Kanno
  • Organizer: The 4th International Congress on Arsenic in the Environment As2012
  • Place of Presentation: Cairns, Australia
• [Presentation] PML発現細胞における亜ヒ酸の影響 (2012)
  • Author(s): 平野靖史郎, 小林弥生, 渡辺喬之
  • Organizer: 第18回ヒ素シンポジウム
  • Place of Presentation: 宮崎, 11月
• [Presentation] PML発現細胞における亜ヒ酸の影響 (2012)
  • Author(s): 平野靖史郎
  • Organizer: 第18回ヒ素シンポジウム
  • Place of Presentation: 宮崎
• [Presentation] 細胞におけるヒ素の代謝と動態 (2012)
  • Author(s): 平野靖史郎
  • Organizer: 第23回日本微量元素学会
  • Place of Presentation: 東京
  • Invited
• [Presentation] Effects of arsenite on macropinocytotic and autophagic activities in CHO-K1 cells. (2012)
  • Author(s): S. Hirano
  • Organizer: The 4th International Congress on Arsenic in the Environment As2012
  • Place of Presentation: ケアンズ
• [Presentation] CHO-K1 細胞における亜ヒ酸のマクロピノサイトーシス阻害効果 (2011)
  • Author(s): 平野靖史郎, 小林弥生, 渡辺喬之, 太田悠葵
  • Organizer: 第16回ヒ素シンポジウム
  • Place of Presentation: 旭川
• [Presentation] Distribution and biliary excretion of arsenic in rats orally administered with dipheneylarsinic acid (2011)
  • Author(s): Kobayashi Y., Yamashiro A., and Hirano S
  • Organizer: 3rd International Symposium on Metallomics
  • Place of Presentation: Muenster, Germany
• [Presentation] Distribution and excretion of arsenicals following a single oral administration of organic arsenic compounds in rats (2011)
  • Author(s): Kobayashi Y., Mizumura A., Yamashiro A, and Hirano S
  • Organizer: 4th International IUPAC Symposium for Trace Elements in Food
  • Place of Presentation: Aberdeen, UK
• [Book] In vivo Testing of Nanomaterials. In “Trend Efficient Designing of Safe Nanomaterials.” (Ed. Jerzy Leszczynski & Tomasz Puzyn) (2013)
  • Author(s): S.Hirano
  • Publisher: RCS Publishing, Royal Society of Chmistry
• [Book] Western blot analysis. Methods Mol Biol, In Nonotoxicology : Methods and Protocol (2012)
  • Author(s): Hirano, Joshua L. Reineke
  • Publisher: Humana Press
• [Book] Western blot analysis in nanotoxicology studies. In “Nonotoxicology: Methods and Protocol (Ed. Joshua L. Reineke) (2012)
  • Author(s): S.Hirano
  • Total Pages: 426
  • Publisher: Humana Press.
• [Remarks] 平野靖史郎のホームページ
  • URL: http://www.nies.go.jp/risk//members/hirano/nano/Self-introduction.html