Heme sensing of the ChrS histidine kinase from Corynebacterium diphtheriae

• Project/Area Number: 21570153
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional biochemistry
• Research Institution: The Institute of Physical and Chemical Research
• Principal Investigator: NAKAMURA Hiroo 独立行政法人理化学研究所, 城生体金属科学研究室, 専任研究員 (80270594)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2011: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2010: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2009: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000)
• Keywords: 病原菌 / ヘム / センサー / 膜タンパク質 / 情報伝達系 / リポソーム / ナノディスク / リン酸化 / 細胞膜 / リン脂質 / 膜タンパク / 鉄 / リン酸化酵素 / 二成分情報伝達系 / 再構成

Research Abstract

It has been hitherto believed that Corynebacterium diphteriae conducts the sensor kinase, ChrS, and the response regulator, ChrA, of a two-component signal transduction system to utilize host heme iron. Although ChrS is predicted to encode a heme sensor, the sensing mechanism remains uncharacterized. ChrS expressed in Eshcherichia coli membranes was solubilized and purified in the presence of a non-ionic detergent. ChrS protein incorporated into proteoliposomes and nanodiscs catalyzed heme-dependent autophosphorylation by ATP. Other metalloporphyrins and iron did not stimulate kinase activity, suggesting that ChrS is a specific heme sensor. This is the first functional reconstitution of a bacterial heme-sensing protein.

Research Products

• [Journal Article] Structure of PAS-linked histidine kinase and the response regulator complex (2009)
  • Author(s): Yamada, S., Sugimoto, H., Kobayashi, M., Ohno, A., Nakamura, H., and Shiro, Y
  • Journal Title: Structure Volume: 17 Pages: 1333-1344
  • Peer Reviewed
• [Journal Article] Heme-dependent autophosphorylation of a heme sensor kinase, ChrS, from Corynebacterium diphtheriae reconstituted in proteoliposomes (2009)
  • Author(s): Ito, Y., Nakagawa, S., Komagata, A., Ikeda-Saito, M., Shiro, Y., and Nakamura, H
  • Journal Title: FEBS Lett Volume: 583 Pages: 2244-2248
  • Peer Reviewed
• [Journal Article] Structure of PAS-linked histidine kinase and the response regulator complex (2009)
  • Author(s): Yamada, S., Sugimoto, H., Kobayashi, M., Ohno, A., Nakamura, H., Shiro, Y.
  • Journal Title: Structure 17 Pages: 1333-1344
  • Peer Reviewed
• [Journal Article] Heme-dependent autophosphorylation of a heme sensor kinase, ChrS, from Corynebacterium diphtheriae reconstituted in proteoliposomes (2009)
  • Author(s): Ito, Y., Nakagawa, S., Komagata, A., Ikeda-Saito, M., Shiro, Y., Nakamura, H.
  • Journal Title: FEBS Lett. 583 Pages: 2244-2248
  • Peer Reviewed
• [Presentation] ナノディスクを用いた病原菌の生体鉄感知システムの解析 (2010)
  • Author(s): 伊藤陽子、當舎武彦、中村寛夫、城宜嗣
  • Organizer: BMB2010第33回日本分子生物学界年会第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-10
• [Presentation] ナノディスクを用いた病原菌の生体鉄感知システムの解析 (2010)
  • Author(s): 伊藤陽子、當舎武彦、中村寛夫、城宜嗣
  • Organizer: 第33回日本分子生物学会年会、第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸、日本
• [Presentation] Heme signal transduction system, ChrS-ChrA, in Corynebacterium diphtheriae : Mutational analysis using the hmuO reporter assay in E. coli (2010)
  • Author(s): Nakamura H
  • Organizer: Gordon Research Conferences on Sensory Transduction in Microorganisms
  • Place of Presentation: Ventura, California, USA
• [Presentation] Heme signal transduction system, ChrS-ChrA, in Corynebacterium diphtheriae : Mutational analysis using the hmuO reporter assay in E.coli (2010)
  • Author(s): 中村寛夫
  • Organizer: Gordon Research Conferences on Sensory Transduction in Microorganisms
  • Place of Presentation: Ventura, California, USA
• [Presentation] ジフテリア菌ヘムセンシングの大腸菌での再構築と機能変異体スクリーニング (2009)
  • Author(s): 益田準一、大柳明日香、城宜嗣、中村寛夫
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜市
  • Year and Date: 2009-12-12
• [Presentation] 脂質ナノディスクに再構成したジフテリア菌ChrSのヘム結合とリン酸基転移 (2009)
  • Author(s): 伊藤陽子、中村寛夫、城宜嗣
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸市
  • Year and Date: 2009-10-22
• [Presentation] ジフテリア菌ヘムセンシングの大腸菌での再構築と機能変異体スクリーニング (2009)
  • Author(s): 益田準一、大柳明日香、城宜嗣、中村寛夫
  • Organizer: 第32回日本分子生物学会年会
  • Place of Presentation: 横浜、日本
• [Presentation] 脂質ナノディスクに再構成したジフテリア菌ChrSのヘム結合とリン酸基転移 (2009)
  • Author(s): 伊藤陽子、中村寛夫、城宜嗣
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸、日本
• [Book] 生体の酸素センサー:細胞レベルの酸素センシング、からだと酸素の事典、酸素ダイナミクス研究会編pp182-189共著 (2009)
  • Author(s): 中村寛夫
  • Total Pages: 596
  • Publisher: 朝倉書店