Molecular structure and functional regulation of ATP-binding cassette transporters

• Project/Area Number: 13142208
• Research Category: Grant-in-Aid for Scientific Research on Priority Areas
• Allocation Type: Single-year Grants
• Review Section: Biological Sciences
• Research Institution: Kyushu University
• Principal Investigator: WADA Morimasa Kyushu University, Department of Medical Biochemistry, Associate Proffesor, 大学院医学研究院, 助教授 (20220965)
• Project Period (FY): 2001 – 2005
• Project Status: Completed (Fiscal Year 2005)
• Budget Amount: ¥70,300,000 (Direct Cost: ¥70,300,000); Fiscal Year 2005: ¥11,000,000 (Direct Cost: ¥11,000,000); Fiscal Year 2004: ¥11,500,000 (Direct Cost: ¥11,500,000); Fiscal Year 2003: ¥17,000,000 (Direct Cost: ¥17,000,000); Fiscal Year 2002: ¥12,800,000 (Direct Cost: ¥12,800,000); Fiscal Year 2001: ¥18,000,000 (Direct Cost: ¥18,000,000)
• Keywords: ABC membrane protein / Effulux pump / Chimeric proteins / Substrate affinity / Colorectal cancer risk / ABCC2 / ABCB1 / 基質親和性 / 大量精製 / 膜ベシクル輸送解析 / 遺伝性黄疸 / Dubin-Johnson症候群 / 発がん / 基質認識 / ABCトランスポーター / MRP2 / ナノマシーン / 機能制御 / 細胞内局在 / ロイコトリエンC4 / 原子構造

Research Abstract

(1) Mechanism of substrate recognition : By constructing of ABCC1/ABCC2 chimeric proteins and examining their the kinetic properties for LTC_4 transport in inside-out membrane vesicles, we have obtained the results suggesting that Membrane Spanning Domain (MSD)1 are involved in substrate affinity and that the COOH-terminal half is exchangeable between two proteins. Because, basic and/or charged residues in MSD2 and MSD3 but not in MSD1 have been reported repeatedly to be involved in substrate binding, our results shows that portion contributing in substrate affinity would be different from that of substrate binding itself.
(2) Sorting mechanisms for intracellular localization : We narrowed down the region participating in determination of apical-basolateral localization to the second membrane spanning domain 2(MSD2)and surrounding region.
(3) To evaluate the effect of ABCB1 in intestinal tumorigenesis directly, we generated Abcbla-disrupted mice with an Apc^<min/+> background, an animal model for human familial adenomatous polyposis, and examined whether polyp formation were affected by the absence of functional ABCB1. The number and size of intestinal polyps in Abcbla^<-/->, Apc^<min/+> mice was significantly smaller than the number in Abcbla^<+/+>, Apc^<Min/+> mice. We then analyzed association between genetic variations in the ABCBI gene and cororectal cancer risk. By case-control study using 460 colorectal cancer patients and 783 healthy volunteers, we obtained the results indicating that the cancer risk of individuals who's genetic background is associated with low ABCB1 level is half of the individuals with lower ABCB1 genetic background.

Research Products

• [Journal Article] Single nucleotide polymorphisms in ABCC2 and ABCB1 genes and their clinical impact in physiology and drug response. (2006)
  • Author(s): Wada, M.
  • Journal Title: Cancer Letters 234 Pages: 40-50
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Single nucleotide polymorphisms in ABCC2 and ABCB1 genes and their clinical impact in physiology and drug response. (2006)
  • Author(s): Wada, M.
  • Journal Title: Cancer Letters (in press)
• [Journal Article] The genetic polymorphism of drug transporters : functional analysis approaches. (2004)
  • Author(s): Ishikawa, T.
  • Journal Title: Pharmacogenomics 5 Pages: 1-33
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Dubin-Johnson-like black liver with normal bilirubin level. (2004)
  • Author(s): Kobayashi, Y.
  • Journal Title: Journal of Gastroenterology 39 Pages: 892-895
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The genetic polymorphism of drug transporters : functional analysis approaches. (2004)
  • Author(s): Ishikawa, T., Tsuji, A., Inui, K., Sai, Y., Anzai, N., Wada, M., Endou, H., Sumino, Y.
  • Journal Title: Pharmacogenomics 5 Pages: 1-33
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Dubin-Johnson-like black liver with normal bilirubin level. (2004)
  • Author(s): Kobayashi, Y., Ishihara, T., Wada, M., Kajihara, S., Araki, J., Mifuji, R., Itani, T., Kuroda, M., Urawa, F., Kaito, M., Adachi, Y.
  • Journal Title: Journal of Gastroenterology 39 Pages: 892-895
  • NAID: 10013613442
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The genetic polymorphism of drug transporters : functional analysis approaches. (2004)
  • Author(s): Ishikawa, T., Wada, M., et al.
  • Journal Title: Pharmacogenomics 5 Pages: 1-33
• [Journal Article] Dubin-Johnson-like black liver with normal bilirubin level. (2004)
  • Author(s): Kobayashi, Y., Wada, M., et al.
  • Journal Title: J.Gastroenterol. 39 Pages: 892-895
  • NAID: 10013613442
• [Journal Article] Genetic polymorphism at the 5' regulatory region of MDR1 and its association with inter-individual variation of expression level in colon. (2003)
  • Author(s): Taniguchi, S.
  • Journal Title: Molecular Cancer Therapeutics 2 Pages: 1351-1359
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The role of P-glycoprotein in intestinal tumorigenesis : disruption of mdrla suppresses polyp formation in Apc^<Min/+> mice. (2003)
  • Author(s): Mochida, Y.
  • Journal Title: Carcinogenesis 24 Pages: 1219-1224
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Identification of Domains Participating in the Substrate Specificity of the Multidrug Resistance Proteins, MRP1 and MRP2. (2003)
  • Author(s): Konno, T.
  • Journal Title: Journal of Biological Chemistry 278 Pages: 22908-22917
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The Basic and Clinical Implication of ABC transporters, YB-1 and Angiogenesis in Human Malignancies. (2003)
  • Author(s): Kuwano, M.
  • Journal Title: Cancer Science 94 Pages: 9-14
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Genetic polymorphism at the 5' regulatory region of MDR1 and its association with inter-individual variation of expression level in colon. (2003)
  • Author(s): Taniguchi, S., Mochida.Y., Uchiumi, T., Tahira, T., Hayashi, K., Takagi, K., Shimada, M., Maehara, Y., Kuwano, H., Kono, S., Nakano, H., Kuwano, M., Wada, M.
  • Journal Title: Molecular Cancer Therapeutics 2 Pages: 1351-1359
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The role of P-glycoprotein in intestinal tumorigenesis : disruption of mdrla suppresses polyp formation in ApcMin/+ mice. (2003)
  • Author(s): Mochida, Y., Taguchi, K., Taniguchi, S., Tsuneyoshi, M., Kuwano, H., Tsuzuki, T., Kuwano, M., Wada, M.
  • Journal Title: Carcinogenesis 24 Pages: 1219-1224
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Identification of Domains Participating in the Substrate Specificity of the Multidrug Resistance Proteins, MRP1 and MRP2. (2003)
  • Author(s): Konno, T., Ebihara, T., Hisaeda, K, Uchiumi, T., Nakamura, T., Shirakusa, T., Kuwano, M., Wada, M.
  • Journal Title: Journal of Biological Chemistry 278 Pages: 22908-22917
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The Basic and Clinical Implication of ABC transporters, YB-1 and Angiogenesis in Human Malignancies. (2003)
  • Author(s): Kuwano, M., Uchiumi, T., Hayakawa, H., Ono, M., Wada, M., Izumi, H., Kohno, K.
  • Journal Title: Cancer Science 94 Pages: 9-14
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Molecular basis of inherited transport defects-, edited by S. Broer and C. A. Wagner : Canalicular multispecific organic anion transporter, ABCC2 (2003)
  • Author(s): Wada, M., Uchiumi, T., Kuwano, M.
  • Journal Title: MEMBRANE TRANSPORT DISEASES -NY KluwerAcadermic/ Plenum Publishers
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Trafficking and functional defects by mutations of the ATP-binding domains in MRP2 in patients with Dubin-Johnson syndrome. (2002)
  • Author(s): Hashimoto, K.
  • Journal Title: Hepatology 36 Pages: 1236-1245
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] The Association of DAP-kinase Hypermethylation with Early Recurrence in Superficial Bladder Cancers (2002)
  • Author(s): Tada, Y.
  • Journal Title: Cancer Research 62 Pages: 4048-4053
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Increased expression of multidrug resistance-associated proteins in bladder cancer during clinical course and drug resistance to doxorubicin. (2002)
  • Author(s): Tada, Y.
  • Journal Title: International Journal of Cancer 98 Pages: 630-635
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Trafficking and functional defects by mutations of the ATP-binding domains in MRP2 in patients with Dubin-Johnson syndrome. (2002)
  • Author(s): Hashimoto, K., Uchiumi, T., Konno, T., Ebihara, T., Nakamura, T., Wada, M., Sakisaka, S., Maniwa, F., Amachi, T., Ueda, K., Kuwano M.
  • Journal Title: Hepatology 36 Pages: 1236-1245
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] The Association of DAP-kinase Hypermethylation with Early Recurrence in Superficial Bladder Cancers (2002)
  • Author(s): Tada, Y., Wada, M., Taguchi, K., Mochida, Y., Kinugawa, N., Tsuneyoshi, M., Naito, S., Kuwano, M.
  • Journal Title: Cancer Research 62 Pages: 4048-4053
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Increased expression of multidrug resistance-associated proteinsin bladder cancer during clinical course and drug resistance to doxorubicin. (2002)
  • Author(s): Tada, Y., Wada, M., Migita, T., Nagayama, J., Hinoshita, E., Mochida, Y., Maehara, Y., Tsuneyoshi, M., Kuwano, M., Naito, S.
  • Journal Title: International Journal of Cancer 98 Pages: 630-635
  • Description: 「研究成果報告書概要(欧文)」より
• [Journal Article] Retrovirus insertion and transcriptional activation of the multidrug resistance (mdrla) gene in leukemias treated by a chemotherapeutic agent in vivo. (2001)
  • Author(s): Nagayama, J.
  • Journal Title: Blood 97 Pages: 759-766
  • Description: 「研究成果報告書概要(和文)」より
• [Journal Article] Retrovirus insertion and transcriptional activation of the multidrug resistance (mdrla) gene in leukemias treated by a chemotherapeutic agent in vivo. (2001)
  • Author(s): Nagayama, J., Iino, M., Tada, Y., Kusaba, H., Kiue, A., Ohshima, K., Kuwano, M., Wada, M.
  • Journal Title: Blood 97 Pages: 759-766
  • Description: 「研究成果報告書概要(欧文)」より
• [Book] MEMBRANE TRANSPORT DISEASES-Molecular basis of inherited transport defects, (edited by S. Broer and C. A. Wagner) : (分担執筆)(Canalicular multispecific organic anion transporter, ABCC2) (2003)
  • Author(s): Wada, M.
  • Total Pages: 390
  • Publisher: NY Kluwer Academic/ Plenum Publishers
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 生命工学、新しい生命へのアプローチ(シリーズ・バイオサイエンスの新世紀) : (分担執筆)酵母人工染色体 (2002)
  • Author(s): 和田守正
  • Total Pages: 298
  • Publisher: 共立出版
  • Description: 「研究成果報告書概要(和文)」より
• [Book] 薬物動態、作用と遺伝子多型 : (分担執筆)MRP2/cMOATの生理機能と変異・遺伝子多型 (2001)
  • Author(s): 和田守正
  • Total Pages: 457
  • Publisher: 医薬ジャーナル社
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] MDR1遺伝子の5'制御領域におけるSNPs (2003)
  • Inventor(s): 和田 守正, 桑野 信彦
  • Industrial Property Rights Holder: 和田 守正, 桑野 信彦
  • Industrial Property Number: 2003-332584
  • Filing Date: 2003-09-24
  • Description: 「研究成果報告書概要(和文)」より
• [Patent(Industrial Property Rights)] ヒトcMOAT遺伝子とそのcDNA (1996)
  • Inventor(s): 和田 守正, 桑野 信彦
  • Industrial Property Rights Holder: 和田 守正, 桑野 信彦
  • Filing Date: 1996-09-30
  • Acquisition Date: 2005-07-08
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Taniguchi, S.他12名: "Genetic polymorphism at the 5' regulatory region of MDR1 and its association with inter-individual variation of expression level in colon."Molecular Cancer Therapeutics. 2. 1351-1359 (2003)
• [Publications] Mochida, Y.他7名: "The role of P-glycoprotein in intestinal tumorigenesis : disruption of mdr1a suppresses polyp formation in Apc^<Min/+> mice."Carcinogenesis. 24. 1219-1224 (2003)
• [Publications] Konno, T.他7名: "Identification of Domains Participating in the Substrate Specificity of the Multidrug Resistance Proteins, MRP1 and MRP2."Journal of Biological Chemistry. 278. 22908-22917 (2003)
• [Publications] Tada, Y., Wada, M.他: "The Association of DAP-kinase Hypermethylation with Early Recurrence in Superficial Bladder Cancers"Cancer Research. 62. 4048-4053 (2002)
• [Publications] Tada, Y., Wada, M.他: "Increased expression of multidrug resistance-associated proteins in bladder cancer during clinical course and drug resistance to doxorubicin"International Journal of Cancer. 98. 630-635 (2002)
• [Publications] Nagayama, J. et al.: "Retrovirus insertion and transcriptional activation of the multidrug resistance(mdr1a) gene in leukemias treated by a chemotherapeutic agent in vivo"Blood. 97. 759-766 (2001)
• [Publications] Tada, Y. et al.: "MDR1 gene overexpression and altered degree of methylation at the promoter region in bladder cancer during chemotherapeutic treatment"Clinical Cancer Research. 6. 4618-4627 (2000)
• [Publications] Harada, T. et al.: "The specific expression of three novel splice variant forms of human metalloprotease-like disintegrin-like cystein-rich protein 2 gene in brain tissues and gliomas"Japanese Journal of Cancer Reseach. 91. 1-6 (2000)
• [Publications] Harada, T. et al.: "Alu-associated interstitial deletions and chromosomal rearrangement in two human multidrug resistant cell lines"International Journal of Cancer. 86. 506-511 (2000)