The elucidation of the zinc supplying mechanism of zinc requiring enzyme by zinc transporters

• Project/Area Number: 22890237
• Research Category: Grant-in-Aid for Research Activity Start-up
• Allocation Type: Single-year Grants
• Research Field: General medical chemistry
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: FUKUNAKA Ayako 独立行政法人理化学研究所, 複数分子イメージング研究チーム, 研究員 (60586402)
• Project Period (FY): 2010 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥3,146,000 (Direct Cost: ¥2,420,000、Indirect Cost: ¥726,000); Fiscal Year 2011: ¥1,508,000 (Direct Cost: ¥1,160,000、Indirect Cost: ¥348,000); Fiscal Year 2010: ¥1,638,000 (Direct Cost: ¥1,260,000、Indirect Cost: ¥378,000)
• Keywords: 亜鉛トランスポーター / 亜鉛要求性酵素 / 分泌経路 / 活性化機構

Research Abstract

The reduced activity of TNAP in DT40 cells deficient of two ZnT complexes(ZnT5/ZnT6 heterodimer and ZnT7 homo-oligomer) was not restored by zinc supplementation nor by exogenous expression of other ZnTs that increase the zinc content in the early secretory pathway. Moreover the expression of ZnT5/ZnT6 heterodimers reconstituted with zinc-transport-incompetent ZnT5 mutant failed to restore TNAP activity, but could stabilize the TNAP protein as the apo-form. These findings demonstrate that TNAP is activated not simply by passive zinc binding, but by an elaborate two-step mechanism via protein stabilization followed by enzyme conversion from the apo-to the holo-form with zinc loaded by ZnT complexes in the early secretory pathway.

Research Products

• [Journal Article] Tissue non-specific alkaline phosphatase is activated via a two-step mechanism by zinc transporter 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 Volume: 286 Pages: 16363-16373
• [Journal Article] Tissue non-specific alkaline phosphatase is activated via a two-step mechanism by zinc transporter 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: Journal of Biological Chemistry Volume: 286 Pages: 16363-16373
  • Peer Reviewed
• [Presentation] 亜鉛トランスポーター複合体による亜鉛要求性酵素の活性化機構の解明 (2011)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 京都
  • Year and Date: 2011-09-24
• [Presentation] 亜鉛トランスポーター複合体による亜鉛要求性酵素の活性化機構の解明 (2011)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都)
  • Year and Date: 2011-09-24
• [Presentation] ZnT transporters activate tissue non-specific alkaline phosphatase by two-step mechanism in the early secretory pathway (2011)
  • Author(s): Ayako Fukunaka, Yayoi Kurokawa, Makoto Hiromura, Shuuichi Enomoto, Taiho Kambe
  • Organizer: The 5^<th> international Conference on Metal and Genetics
  • Place of Presentation: Kobe
  • Year and Date: 2011-09-06
• [Presentation] ZnT transporters activate tissue non-specific alkaline phosphatase by two-step mechanism in the early secretory pathway (2011)
  • Author(s): Ayako Fukunaka, Yayoi Kurokawa, Makoto Hiromura, Shuuichi Enomoto, Taiho Kambe
  • Organizer: The 5^<th> international Conference on Metal and Genetics
  • Place of Presentation: Kobe International Conference Center(神戸)
  • Year and Date: 2011-09-06
• [Presentation] 亜鉛トランスポーター複合体による亜鉛要求性酵素の活性化機構の解明 (2011)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第22回日本微量金属元素学会
  • Place of Presentation: 京都
  • Year and Date: 2011-07-01
• [Presentation] 亜鉛トランスポーター複合体による亜鉛要求性酵素の活性化機構の解明 (2011)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第22回日本微量金属元素学会
  • Place of Presentation: 京都テルサ/京都府民総合交流プラザ(京都)
  • Year and Date: 2011-07-01
• [Presentation] Demonstration and characterization of th e heterodimerization of ZnT5 and ZnT6 in the early secretory pathway (2010)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第2回メタロミクス研究フォーラム
  • Place of Presentation: 京都
  • Year and Date: 2010-11-03
• [Presentation] Demonstration and characterization of the heterodimerization of ZnT5 and ZnT6 in the early secretory pathway (2010)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第2回メタロミクス研究フォーラム
  • Place of Presentation: 京都薬科大学 愛学館 (京都市)
  • Year and Date: 2010-11-03
• [Presentation] Demonstration and characterization of th e heterodimerization of ZnT5 and ZnT6 in the early secretory pathway (2010)
  • Author(s): Ayako Fukunaka, Yayoi Kurokawa, Makoto Hiromura, Syuuichi Enomoto, Taiho Kambe
  • Organizer: The 60^<th> Fujihara Seminar Zinc signaling and cellular functions
  • Place of Presentation: Osaka
  • Year and Date: 2010-10-29
• [Presentation] Demonstration and characterization of the heterodimerization of ZnT5 and ZnT6 in the early secretory pathway (2010)
  • Author(s): Ayako Fukunaka, Yayoi Kurokawa, Makoto Hiromura, Syuuichi Enomoto, Taiho Kambe
  • Organizer: The 60^<th> Fujihara Seminar Zinc signaling and cellular functions
  • Place of Presentation: Grand Cube Osaka (大阪市)
  • Year and Date: 2010-10-29
• [Presentation] 亜鉛トランスポーターZnT5/ZnT6ヘテロ複合体の性状解析 (2010)
  • Author(s): 福中彩子、黒川弥生、廣村信、榎本秀一、神戸大朋
  • Organizer: 第21回日本微量金属元素
  • Place of Presentation: 京都
  • Year and Date: 2010-07-04
• [Presentation] Conversion of the apoenzymes to the zinc-containing holoenzymes by ZnT transporters (2010)
  • Author(s): 福中彩子、神戸大朋
  • Organizer: 第20回金属の関与する生体関連反応シンポジウム
  • Place of Presentation: 徳島
  • Year and Date: 2010-06-25
• [Book] 亜鉛酵素の活性化メカニズムを解明,理研の最前線
  • Author(s): 福中彩子
  • Publisher: 日刊工業新聞
• [Remarks] プレスリリース
  • URL: http://www.riken.jp/r-world/research/results/2011/110426/index.html
• [Remarks]
  • URL: http://www.rikenresearch.riken.jp/eng/research/6654
• [Remarks]
  • URL: http://www.riken.jp/r-world/research/results/2011/110426/index.html
• [Remarks]
  • URL: http://www.rikenresearch.riken.jp/eng/research/6654