Identification of sub-femto mole protein from gel electrophoresis

• Project/Area Number: 19590275
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Kyoto University
• Principal Investigator: OKAWA Katsuya Kyoto University, 医学研究科, その他(グループリーダー) (20444458)
• Project Period (FY): 2007 – 2008
• Project Status: Completed (Fiscal Year 2008)
• Budget Amount: ¥4,550,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥1,050,000); Fiscal Year 2008: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2007: ¥2,860,000 (Direct Cost: ¥2,200,000、Indirect Cost: ¥660,000)
• Keywords: 生体分子医学 / 蛋白質 / プロテオーム / 分析科学 / 電気泳動 / サブフェムトモル

Research Abstract

従来法の要素技術の検証を進め、改善点が見出されて来ている.これまでに開発した、従来の10倍程度以上高感度なゲル内タンパク質染色法を活用し、本方法でなければ可視化出来ないサブフェムトモルのモデルとなるタンパク質を調製し、少なくともサブフェムトモルレベルでは従来法の延長線上の解析手法にて対応可能であることが見出された.
また、得られた検証結果を随時、実践のタンパク質解析に応用している.

Research Products

• [Journal Article] mDia2 shuttles between the nucleus and the cytoplasm through the Importin-{alpha}/{beta}- and CRM1-mediated nuclear transport mechanism (2009)
  • Author(s): Miki T, Okawa K, Sekimoto T, Yoneda Y, Watanabe S, Ishizaki T, Narumiya S
  • Journal Title: J Biol Chem 284(9) Pages: 5753-62
  • Peer Reviewed
• [Journal Article] Isolation and characterization of postsplicing lariat-intron complexes (2009)
  • Author(s): Yoshimoto R, Kataoka N, Okawa K, Ohno M
  • Journal Title: Nucleic Acids Res 37(3) Pages: 891-902
  • Peer Reviewed
• [Journal Article] Anti-moesin antibodies in the serum of patients with aplastic anemia stimulate peripheral blood mononuclear cells to secrete TNF-alpha and IFN-gamma (2009)
  • Author(s): Takamatsu H, Espinoza JL, Lu X, Qi Z, Okawa K, Nakao S
  • Journal Title: J Immunol 182(1) Pages: 703-10
  • Peer Reviewed
• [Journal Article] RECK forms cowbell-shaped dimers and inhibits matrix metalloproteinase-catalyzed cleavage of fibronectin (2009)
  • Author(s): Omura A, Matsuzaki T, Mio K, Ogura T, Yamamoto M, Fujita A, Okawa K, Kitayama H, Takahashi C, Sato C, Noda M
  • Journal Title: J Biol Chem 284(6) Pages: 3461-9
  • Peer Reviewed
• [Journal Article] Isolation and characterization of post-splicing lariat-intron complex (2009)
  • Author(s): Yoshimoto R, ほか
  • Journal Title: Nucleic Acids Res. 37 Pages: 891-902
  • Peer Reviewed
• [Journal Article] CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore (2008)
  • Author(s): Hori T, Amano M, Suzuki A, Backer CB, Welburn JP, Dong Y, McEwen BF, Shang WH, Suzuki E, Okawa K, Cheeseman IM, Fukagawa T
  • Journal Title: Cell 135(6) Pages: 1039-52
  • Peer Reviewed
• [Journal Article] IQGAP3 regulates cell proliferation through the Ras/ERK signalling cascade (2008)
  • Author(s): Nojima H, Adachi M, Matsui T, Okawa K, Tsukita S, Tsukita S
  • Journal Title: Nat Cell Biol 10(8) Pages: 971-8
  • Peer Reviewed
• [Journal Article] Human XTP3-B forms an endoplasmic reticulum quality control scaffold with the HRD1-SEL1L ubiquitin ligase complex and BiP (2008)
  • Author(s): Hosokawa N, Wada I, Nagasawa K, Moriyama T, Okawa K, Nagata K
  • Journal Title: J Biol Chem 283(30) Pages: 20914-24
  • Peer Reviewed
• [Journal Article] Optimized proteomic analysis on gels of cell-cell adhering junctional membrane proteins (2008)
  • Author(s): Yamazaki Y, Okawa K, Yano T, Tsukita S, Tsukita S
  • Journal Title: Biochemistry 47(19) Pages: 5378-86
  • Peer Reviewed
• [Journal Article] Loss of Borealin/DasraB leads to defective cell proliferation, p53 accumulation and early embryonic lethality (2008)
  • Author(s): Yamanaka Y, Heike T, Kumada T, ShibataM, Takaoka Y, Kitano A, Shiraishi K, Kato T, Nagato M, Okawa K, Furushima K, Nakao K, Nakamura Y, Taketo MM, Aizawa S, Nakahata T
  • Journal Title: Mech Dev 125(5-6) Pages: 441-50
  • Peer Reviewed
• [Journal Article] Biochemical characterization of the Rho GTPase-regulated actin assembly by diaphanous-related formins, mDia1 and Daam1, in platelets (2008)
  • Author(s): Higashi T, Ikeda T, Shirakawa R, Kondo H, Kawato M, Horiguchi M, Okuda T, Okawa K, Fukai S, Nureki O, Kita T, Horiuchi H
  • Journal Title: J Biol Chem 283(13) Pages: 8746-55
  • Peer Reviewed
• [Journal Article] MKKS is a centrosome-shuttling protein degraded by disease-causing mutations via CHIP-mediated ubiquitination (2008)
  • Author(s): Hirayama S, Yamazaki Y, Kitamura A, Oda Y, Morito D, Okawa K, Kimura H, Cyr DM, Kubota H, Nagata K.
  • Journal Title: Mol Biol Cell 19(3) Pages: 899-911
  • Peer Reviewed
• [Journal Article] Regulation of platelet dense granule secretion by the Ral GTPase-exocyst pathway (2008)
  • Author(s): Kawato M, Shirakawa R, Kondo H, Higashi T, Ikeda T, Okawa K, Fukai S, Nureki O, Kita T, Horiuchi H
  • Journal Title: J Biol Chem 283(1) Pages: 166-74
  • Peer Reviewed
• [Journal Article] IQGAP3 regulates cell proliferation through the Ras/ERK signalling cascade. (2008)
  • Author(s): Nojima H, ほか
  • Journal Title: Nat Cell Biol. 10 Pages: 971-8
  • Peer Reviewed
• [Journal Article] CCAN makes multiple contacts with centromeric DNA to provide distinct pathways to the outer kinetochore. (2008)
  • Author(s): Hori T, ほか
  • Journal Title: Cell. 135 Pages: 1039-52
  • Peer Reviewed
• [Journal Article] MKKS Is a Centrosome-shuttling Protein Degraded by Disease-causing Mutations via CHIP-mediated Ubiquitination. (2008)
  • Author(s): Hirayama S, ほか
  • Journal Title: Mol Biol Cell 19 Pages: 899-911
  • Peer Reviewed
• [Journal Article] In vitro analyses of the production and activity of secondary small interfering RNAs in C. elegans (2007)
  • Author(s): Aoki K, Moriguchi H, Yoshioka T, Okawa K, Tabara H.
  • Journal Title: EMBO J 26(24) Pages: 5007-19
  • Peer Reviewed
• [Journal Article] Enhancement of alpha-secretase cleavage of amyloid precursor protein by a metalloendopeptidase nardilysin (2007)
  • Author(s): Hiraoka Y, Ohno M, Yoshida K, Okawa K, Tomimoto H, Kita T, Nishi E
  • Journal Title: J Neurochem 102(5) Pages: 1595-605
  • Peer Reviewed
• [Journal Article] Heat-shock protein 105 interacts with and suppresses aggregation of mutant Cu/Zn superoxide dismutase : clues to a possible strategy for treating ALS (2007)
  • Author(s): Yamashita H, Kawamata J, Okawa K, Kanki R, Nakamizo T, Hatayama T, Yamanaka K, Takahashi R, Shimohama S
  • Journal Title: J Neurochem 102(5) Pages: 1497-505
  • Peer Reviewed
• [Journal Article] Factors associated with a purine-rich exonic splicing enhancer sequence in Xenopus oocyte nucleus (2007)
  • Author(s): Masuyama K, Taniguchi I, Okawa K, OhnoM
  • Journal Title: Biochem Biophys Res Commun 359(3) Pages: 580-5
  • Peer Reviewed
• [Journal Article] Control of immune responses by antigen-specific regulatory T cells expressing the folate receptor (2007)
  • Author(s): Yamaguchi T, Hirota K, Nagahama K, Okawa K, Takahashi T, Nomura T, Sakaguchi S
  • Journal Title: Immunity 27(1) Pages: 145-59
  • Peer Reviewed
• [Journal Article] Gab family proteins are essential for postnatal maintenance of cardiac function via neuregulin-1/ErbB signaling (2007)
  • Author(s): Nakaoka Y, Nishida K, Narimatsu M, Kamiya A, Minami T, Sawa H, Okawa K, Fujio Y, Koyama T, Maeda M, Sone M, Yamasaki S, Arai Y, Koh GY, Kodama T, Hirota H, Otsu K, Hirano T, Mochizuki N
  • Journal Title: J Clin Invest 117(7) Pages: 1771-81
  • Peer Reviewed
• [Journal Article] Cyclin-dependent kinaselike 5 is a novel target of immunotherapy in adult T-cell leukemia (2007)
  • Author(s): Kawahara M, Hori T, Matsubara Y, Okawa K, Uchiyama T
  • Journal Title: J Immunother 30(5) Pages: 499-505
  • Peer Reviewed
• [Journal Article] The reversion-inducing cysteine-rich protein with Kazal motifs(RECK)interacts with membrane type 1 matrix metalloproteinase and CD13/aminopeptidase N and modulates their endocytic pathways (2007)
  • Author(s): Miki T, Takegami Y, Okawa K, Muraguchi T, Noda M, Takahashi C
  • Journal Title: J Biol Chem 282(16) Pages: 12341-52
  • Peer Reviewed
• [Journal Article] Gab family proteins are essential for postnatal maintenance of cardiac function via neuregulin-1/ErbB signaling. (2007)
  • Author(s): Nakaoka Y, ほか
  • Journal Title: J Clin Invest 117 Pages: 1771-81
  • Peer Reviewed
• [Journal Article] Control of immune responses by antigen-specific regulatory Tcells expressing the folate receptor. (2007)
  • Author(s): Yamaguchi T, ほか
  • Journal Title: Immunity 27 Pages: 145-59
  • Peer Reviewed
• [Presentation] IQGAP3 regulates cell proliferation through the Ras/ERK signaling cascade (2008)
  • Author(s): H Nojima, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] 新規レクチンによる小胞体タンパク質品質管理機構 (2008)
  • Author(s): 細川暢子, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] Daam1、mDia1によるアクチン重合反応制御機構 (2008)
  • Author(s): 東智仁, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] セントロメアDNAに結合するCENP-T複合体の機能解析 (2008)
  • Author(s): 堀哲也, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] Adaptor Nck1 induces increased tyrosine phosphorylation of GIT2 (2008)
  • Author(s): T Sasaki, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] mRNA切断活性を示す線虫のArgonaute蛋白についての遺伝生化学的解析 (2008)
  • Author(s): 田原浩昭, ほか
  • Organizer: BMB
  • Place of Presentation: 神戸
• [Presentation] TAP-MS法によるインテグレス結合因子Huwe1の同定とその解析ース結合因子Huwe1の同定とその解析 (2008)
  • Author(s): 山元誠司, ほか
  • Organizer: 第22回日本エイズ学会学術集会・総会
  • Place of Presentation: 大阪
• [Presentation] ROLE OF TDRD9 IN MALE MEIOSIS AND REGULATORY PATHWAY OF TRANSPOSON RNA AND DNA METHYLATION (2008)
  • Author(s): M Shoji, ほか
  • Organizer: 6th biennial meeting on Germ Cells
  • Place of Presentation: CSH
• [Presentation] AJ fraction を用いた膜タンパク質のプロテオミクス解析 (2008)
  • Author(s): 山崎裕自, ほか
  • Organizer: 第60回日本細胞生物学会大会
  • Place of Presentation: 横浜
• [Presentation] IDENTIFICATION OF CELLULAR INTERACTORS TO MoMLV INTEGRASE USING TANDEM AFFINITY PURIFICATION - MASS SPECTROMETRY ANALYSIS (2008)
  • Author(s): S Yamamoto, ほか
  • Organizer: 33rd annual meeting on Retroviruses
  • Place of Presentation: CSH
• [Presentation] Rhoによって制御される細胞質中のForminタンパク質によるアクチン重合 (2007)
  • Author(s): 東 智仁, ほか
  • Organizer: BMB2007
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2007-12-12
• [Presentation] 血小板における dynaminの発現-アクチン重合への関与の可能性 (2007)
  • Author(s): 漆原範子, ほか
  • Organizer: BMB
  • Place of Presentation: 横浜
• [Presentation] 小胞体関連分解構成因子Derlinsに結合するタンパク質の同定と解析 (2007)
  • Author(s): 蜷川暁, ほか
  • Organizer: BMB
  • Place of Presentation: 横浜
• [Presentation] Cell adhesion regulates the interaction between Nck and GIT1 (2007)
  • Author(s): 佐々木知幸, ほか
  • Organizer: BMB
  • Place of Presentation: 横浜
• [Presentation] Rhoによって制御される細胞質中のForminタンパク質によるアクチン重合 (2007)
  • Author(s): 東智仁, ほか
  • Organizer: BMB
  • Place of Presentation: 横浜
• [Presentation] C. elegans における二次型siRNAの産生と活性についての遺伝生化学的解析 (2007)
  • Author(s): 田原浩昭, ほか
  • Organizer: BMB
  • Place of Presentation: 横浜
• [Remarks]
  • URL: http://sentan1.hmro.med.kyoto-u.ac.jp/?k-okawa/
• [Remarks]
  • URL: http://sentan1.hmro.med.kyoto-u.ac.jp/~k-okawa/