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Biological strategy for high quality and productivity of recombinant proteins in animal cells by the use of metal transporters

Research Project

Project/Area Number 21780309
Research Category

Grant-in-Aid for Young Scientists (B)

Allocation TypeSingle-year Grants
Research Field Applied molecular and cellular biology
Research InstitutionKyoto University

Principal Investigator

KAMBE Taiho  Kyoto University, 大学院・生命科学研究科, 准教授 (90303875)

Project Period (FY) 2009 – 2010
Project Status Completed (Fiscal Year 2010)
Budget Amount *help
¥4,680,000 (Direct Cost: ¥3,600,000、Indirect Cost: ¥1,080,000)
Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2009: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
Keywords細胞工学 / 物質生産 / 糖鎖 / マンガン / トランスポーター / 組換え型タンパク質 / 動物細胞 / 微量金属 / 組み換え型タンパク質
Research Abstract

Mammalian cells are used for producing recombinant proteins whose post-translational modifications are important for their therapeutic efficacy. The manufacture of recombinant proteins is complicated by the need for both high levels of expression and appropriate processing of the nascent polypeptide, especially in glycoproteins, because the capacity of glycosylation in the secretory pathway is limited in the cells. In this study, we tried to improve the strategy for producing recombinant glycoproteins by identify metal transporters that supply metal such as zinc and manganese to glycosyltransferases in the secretory pathway because these metals are functional as cofactors of the enzymes. We found a homologue of Pmr1 that is a functional as calcium cannel in yeast is important for manganese transport and likely functional in glycosylation reaction in the secretory pathway, which may lead to the improvement of the secretion capacities of recombinant cells.

Report

(3 results)
  • 2010 Annual Research Report   Final Research Report ( PDF )
  • 2009 Annual Research Report
  • Research Products

    (45 results)

All 2011 2010 2009 Other

All Journal Article (23 results) (of which Peer Reviewed: 17 results) Presentation (21 results) Remarks (1 results)

  • [Journal Article] Tissue non-specific alkaline phosphatase is activated via a two-step mechanism by zinc transport complexes in the early secretory pathway,2011

    • Author(s)
      Fukunaka A, Kurokawa Y, Teranishi F, Sekler I, Oda K, Ackland M. L, Faundez V, Hiromura M, Masuda S, Nagao M, Enomoto S, Kambe T
    • Journal Title

      J.Biol.Chem. 286

      Pages: 16363-16373

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] 生体機能における亜鉛トランスポーターの重要性,2011

    • Author(s)
      神戸大朋
    • Journal Title

      亜鉛トランスポーターをめぐる最近の知見-亜鉛栄養治療 1

      Pages: 54-64

    • Related Report
      2010 Final Research Report
  • [Journal Article] 分泌経路において機能する亜鉛トランスポーター,-亜鉛酵素の活性化との関わり-」2010

    • Author(s)
      神戸大朋
    • Journal Title

      Biomedical Research on Trace Elements-日本微量元素学会誌 21

      Pages: 25-31

    • NAID

      10027734181

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] 亜鉛トランスポーターZnTとZIPの亜鉛輸送機構2010

    • Author(s)
      福中彩子、神戸大朋
    • Journal Title

      生化学 82

      Pages: 30-34

    • NAID

      10026300213

    • Related Report
      2010 Final Research Report
  • [Journal Article] 分泌経路において機能する亜鉛トランスポーター-亜鉛酵素の活性化との関わり-2010

    • Author(s)
      神戸大朋
    • Journal Title

      Biomedical Research on Trace Elements-日本微量元素学会誌

      Volume: 21 Pages: 25-31

    • NAID

      10027734181

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 亜鉛トランズポーターZnTとZIPの亜鉛輸送機構2010

    • Author(s)
      福中彩子、神戸大朋
    • Journal Title

      生化学

      Volume: 82 Pages: 30-34

    • Related Report
      2010 Annual Research Report
  • [Journal Article] Demonstration and characterization of the hetero-dimerization of ZnT5 and ZnT6 in the early secretory pathway2009

    • Author(s)
      Fukunaka A, Suzuki T, Kurokawa Y, Yamazaki T, Fujiwara N, Ishihara K, Migaki H, Okumura K, Masuda S, Yamaguchi-Iwai Y, Nagao M, Kambe T
    • Journal Title

      J.Biol.Chem. 284

      Pages: 30798-30806

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Fritsche K, and Petris M. J, A roll for the ATP7A copper-transporting ATPase in macrophage bactericidal activity2009

    • Author(s)
      White C, Lee J, Kambe T, Fritsche K, Petris M.
    • Journal Title

      J.Biol.Chem. 284

      Pages: 33949-33956

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Yifrach O, Hershfinkel M and Sekler I, Identification of the Zn2+ binding site and mode of operation of a mammalian Zn2+ transporter2009

    • Author(s)
      Ohana E, Hoch E, Keisar C, Kambe T
    • Journal Title

      J.Biol.Chem. 284

      Pages: 17677-17686

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] SLC39A9 (ZIP9) regulates zinc homeostasis in the secretory pathway : haracterization of the ZIP subfamily I protein in vertebrate cells2009

    • Author(s)
      Matsuura W, Yamazaki T, Yamaguchi-Iwai Y, Masuda S, Nagao M, Andrews G.K, Kambe T
    • Journal Title

      Biosci.Biotechnol. Biochem. 73

      Pages: 1142-1148

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Fulcher Y. G, Sachdev S. W, Bush A. I, Fritsche K, Lee J, Quinn T. P, and Petris M.J, "Copper transport into the secretory pathway is regulated by oxygen in macrophages2009

    • Author(s)
      White C, Kambe T
    • Journal Title

      J.Cell.Sci. 122

      Pages: 1315-1321

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Novel proteolytic processing of the ectodomain of the zinc transporter ZIP4 (Slc39a4) during zinc deficiency is inhibited by acrodermatitis enteropathica mutations2009

    • Author(s)
      Kambe T, Andrews G.K
    • Journal Title

      Mol Cell Biol. 29

      Pages: 129-139

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] 亜鉛トランスポーター研究から見た亜鉛バイオロジー2009

    • Author(s)
      神戸大朋
    • Journal Title

      Trace Nutrients Research-微量栄養素研究- 26

      Pages: 11-16

    • NAID

      40017002247

    • Related Report
      2010 Final Research Report 2009 Annual Research Report
  • [Journal Article] 亜鉛の生理機能を司る亜鉛トランスポーター2009

    • Author(s)
      神戸大朋
    • Journal Title

      化学と生物 47

      Pages: 545-552

    • NAID

      10026436172

    • Related Report
      2010 Final Research Report 2009 Annual Research Report
  • [Journal Article] Demonstration and characterization of the hetero-dimerization of ZnT5 and ZnT6 in the early secretory pathway2009

    • Author(s)
      Fukunaka A, Suzuki T, Kurokawa Y, Yamazaki T, Fujiwara N, Ishihara K, Migaki H, Okumura K, Masuda S, Yamaguchi-Iwai Y, Nagao M, Kambe T
    • Journal Title

      The Journal of Biological Chemistry 284

      Pages: 30798-30806

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Identification of the Zn2+ binding site and mode of operation of a mammalian Zn2+ transporter2009

    • Author(s)
      Ohana E, Hoch E, Keisar C, Kambe T, Yifrach O, Hershfinkel M, Sekler I
    • Journal Title

      The Journal of Biological Chemistry 284

      Pages: 17677-17686

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] SLC39A9 (ZIP9) regulates zinc homeostasis in the secretory pathway : Characterization of the ZIP subfamily I protein in vertebrate cells2009

    • Author(s)
      Matsuura W, Yamazaki T, Yamaguchi-Iwai Y, Masuda S, Nagao M, Andrews G.K, Kambe T
    • Journal Title

      Biosci.Biotechnol.Biochem. 73

      Pages: 1142-1148

    • Related Report
      2009 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Molecular and genetic features of zinc transporters in physiology and pathogenesis

    • Author(s)
      Fukada T, Kambe T
    • Journal Title

      Metallomics (in press)

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] An Overview of a Wide Range of Functions of ZnT and Zip Zinc Transporters in the Secretory Pathway

    • Author(s)
      Kambe T
    • Journal Title

      Biosci.Biotechnol. Biochem. (in press)

    • Related Report
      2010 Final Research Report
    • Peer Reviewed
  • [Journal Article] Tissue non-specific alkaline phosphatase is activated via a two-step mechanism by zinc transport complexes in the early secretory pathway

    • Author(s)
      Fukunaka A, Kurokawa Y, Teranishi F, Sekler I, Oda K, Ackland M.L, Faundez V, Hiromura M, Masuda S, Nagao M, Enomoto S, Kambe T
    • Journal Title

      The Journal of Biological Chemistry

      Volume: (in press)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] Molecular and genetic features of zinc transporters in physiology and pathogenesis

    • Author(s)
      Fukada T, Kambe T
    • Journal Title

      Metallomics

      Volume: (in press)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] An Overview of a Wide Range of Functions of ZnT and Zip Zinc Transporters in the Secretory Pathway

    • Author(s)
      Kambe T
    • Journal Title

      Biosci.Biotechnol.Biochem.

      Volume: (in press)

    • Related Report
      2010 Annual Research Report
    • Peer Reviewed
  • [Journal Article] 生体機能における亜鉛トランスポーターの重要性-亜鉛トランスポーターをめぐる最近の知見-

    • Author(s)
      神戸大朋
    • Journal Title

      亜鉛栄養治療

      Volume: (印刷中)

    • Related Report
      2010 Annual Research Report
  • [Presentation] 生体機能における亜鉛トランスポーターの重要性-亜鉛トランスポーターをめぐる最近の知見-2011

    • Author(s)
      Taiho Kambe
    • Organizer
      第2回近畿亜鉛栄養治療研究会
    • Place of Presentation
      シノテスト大阪支店(大阪府)
    • Year and Date
      2011-02-05
    • Related Report
      2010 Final Research Report
  • [Presentation] 生体機能における亜鉛トランスポーターの重要性-亜鉛トランスポーターをめぐる最近の知見-2011

    • Author(s)
      神戸大朋
    • Organizer
      第2回近畿亜鉛栄養治療研究会
    • Place of Presentation
      シノテスト大阪支店 (大阪府)
    • Year and Date
      2011-02-05
    • Related Report
      2010 Annual Research Report
  • [Presentation] 分子遺伝学的手法を用いた亜鉛トランスポーターの機能に関する研究2011

    • Author(s)
      神戸大朋
    • Organizer
      農芸化学会関西支部大会
    • Place of Presentation
      楽友会館(京都府)
    • Year and Date
      2011-02-05
    • Related Report
      2010 Annual Research Report
  • [Presentation] 分泌経路における亜鉛トランスポーターの役割2010

    • Author(s)
      神戸大朋
    • Organizer
      北陸合同バイオシンポジウム2010
    • Place of Presentation
      芦泉荘(福井県)
    • Year and Date
      2010-11-13
    • Related Report
      2010 Annual Research Report
  • [Presentation] Roles of zinc transporters in the early secretory pathway2010

    • Author(s)
      Taiho Kambe
    • Organizer
      The 60th Fujihara Seminar : Zinc Signaling and Cellular Functions
    • Place of Presentation
      大阪国際会議場(大阪府)
    • Year and Date
      2010-10-29
    • Related Report
      2010 Final Research Report
  • [Presentation] Roles of zinc transporters in the early secretory pathway2010

    • Author(s)
      Taiho Kambe
    • Organizer
      The 60th Fujihara Seminar : Zinc Signaling and Cellular Functions
    • Place of Presentation
      大阪国際会議場(大阪府)(招待講演)
    • Year and Date
      2010-10-29
    • Related Report
      2010 Annual Research Report
  • [Presentation] 消化管からの亜鉛吸収に機能するZIPトランスポーターの制御機構2010

    • Author(s)
      神戸大朋
    • Organizer
      第21回日本微量元素学会学術集会
    • Place of Presentation
      京都大学百周年時計台記念館(京都府)
    • Year and Date
      2010-07-04
    • Related Report
      2010 Annual Research Report 2010 Final Research Report
  • [Presentation] Homeostatic Maintenance of Secretory Pathway Function by Zinc Transport complexes2010

    • Author(s)
      Taiho Kambe
    • Organizer
      The 1st PT-ERC(ISPM 2010)
    • Place of Presentation
      Cheongju, Korea
    • Year and Date
      2010-06-26
    • Related Report
      2010 Final Research Report
  • [Presentation] Homeostatic Maintenance of Secretory Pathway Function by Zinc Transport complexes2010

    • Author(s)
      Taiho Kambe
    • Organizer
      The 1st PT-ERC(ISPM 2010)
    • Place of Presentation
      Cheongju, Korea(招待講演)
    • Year and Date
      2010-06-26
    • Related Report
      2010 Annual Research Report
  • [Presentation] 分泌型亜鉛酵素のポロ酵素への変換と亜鉛トランスポーター2010

    • Author(s)
      神戸大朋
    • Organizer
      第10回日本蛋白質科学会シンポジウム
    • Place of Presentation
      札幌コンベンションセンター(北海道)
    • Year and Date
      2010-06-17
    • Related Report
      2010 Final Research Report
  • [Presentation] 分泌型亜鉛酵素のホロ酵素への変換と亜鉛トランスポーター2010

    • Author(s)
      神戸大朋
    • Organizer
      第10回日本蛋白質科学会シンポジウム
    • Place of Presentation
      札幌コンベンションセンター(北海道)
    • Year and Date
      2010-06-17
    • Related Report
      2010 Annual Research Report
  • [Presentation] Regulation of mammalian SLC39A transporters-Zinc-regulated processing of mZIP4-2010

    • Author(s)
      Taiho Kambe
    • Organizer
      Japan/Korea Joint Symposium on Life Science
    • Place of Presentation
      Seoul, Korea
    • Year and Date
      2010-05-28
    • Related Report
      2010 Final Research Report
  • [Presentation] Regulation of mammalian SLC39A transporters-Zinc-regulated processing of mZIP4-2010

    • Author(s)
      Taiho Kambe
    • Organizer
      Japan/Korea Joint Symposium on Life Science
    • Place of Presentation
      Seoul, Korea(招待講演)
    • Year and Date
      2010-05-28
    • Related Report
      2010 Annual Research Report
  • [Presentation] 分子遺伝学的手法を用いた亜鉛トランスポーターの機能に関する研究2010

    • Author(s)
      神戸大朋
    • Organizer
      日本農芸化学会2010年度大会
    • Place of Presentation
      東京
    • Year and Date
      2010-03-27
    • Related Report
      2009 Annual Research Report
  • [Presentation] ZIPとZnT-亜鉛の生理機能を統御する亜鉛トランスポーター-2010

    • Author(s)
      神戸大朋
    • Organizer
      北陸大学学術フロンティア講演会
    • Place of Presentation
      金沢
    • Year and Date
      2010-01-29
    • Related Report
      2009 Annual Research Report
  • [Presentation] Regulation of mammalian SLC39A transporters-Zinc-regulated processing of mZIP42009

    • Author(s)
      Taiho Kambe, Glen K.Andrews
    • Organizer
      International society for Zinc Biology 2009 meeting
    • Place of Presentation
      イスラエル(エルサレム)
    • Year and Date
      2009-12-05
    • Related Report
      2010 Final Research Report
  • [Presentation] Regulation of mammalian SLC39A transporters -Zinc-regulated processing of mZIP42009

    • Author(s)
      Taiho Kambem, Glen K.Andrews
    • Organizer
      International society for Zinc Biology 2009 meeting
    • Place of Presentation
      イスラエル(エルサレム)
    • Year and Date
      2009-12-05
    • Related Report
      2009 Annual Research Report
  • [Presentation] 生体微量金属ホメオスタシスとその輸送システム2009

    • Author(s)
      神戸大朋
    • Organizer
      第82回日本生化学会大会シンポジウム
    • Place of Presentation
      神戸
    • Year and Date
      2009-10-23
    • Related Report
      2010 Final Research Report 2009 Annual Research Report
  • [Presentation] 亜鉛トランスポーター研究から見た亜鉛バイオロジー2009

    • Author(s)
      神戸大朋
    • Organizer
      メタルチオネインおよびメタルバイオサイエンス研究会
    • Place of Presentation
      東京
    • Year and Date
      2009-10-16
    • Related Report
      2009 Annual Research Report
  • [Presentation] 分泌経路における亜鉛酵素の活性化機構-亜鉛トランスポーターによる制御-2009

    • Author(s)
      神戸大朋
    • Organizer
      第20回日本微量元素学会学術集会
    • Place of Presentation
      東京
    • Year and Date
      2009-07-03
    • Related Report
      2009 Annual Research Report
  • [Presentation] 亜鉛トランスポーター研究から始まった亜鉛バイオロジーの新たな展開2009

    • Author(s)
      神戸大朋
    • Organizer
      第26回日本微量栄養素学会学術集会
    • Place of Presentation
      京都
    • Year and Date
      2009-05-30
    • Related Report
      2009 Annual Research Report
  • [Remarks] ホームページ等

    • URL

      http://www.lif.kyoto-u.ac.jp/labs/seitaijoho/

    • Related Report
      2010 Final Research Report

URL: 

Published: 2009-04-01   Modified: 2016-04-21  

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