リン酸化とユビキチン化のクロストークによる腎糸球体タンパク質分解の網羅的解析

• Project/Area Number: 14F04105
• Research Category: Grant-in-Aid for JSPS Fellows
• Allocation Type: Single-year Grants
• Section: 外国
• Research Field: Kidney internal medicine
• Research Institution: Niigata University
• Principal Investigator: 山本 格 新潟大学, 産学地域連携推進機構, 教授 (30092737)
• Co-Investigator(Kenkyū-buntansha): ABDELGAWWAD SAMEH 新潟大学, 産学地域連携推進機構, 外国人特別研究員 ; ABDELGAWWAD Sameh 新潟大学, 医歯学系, 外国人特別研究員
• Project Period (FY): 2014-04-25 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥2,300,000 (Direct Cost: ¥2,300,000); Fiscal Year 2015: ¥1,100,000 (Direct Cost: ¥1,100,000); Fiscal Year 2014: ¥1,200,000 (Direct Cost: ¥1,200,000)
• Keywords: プロテオーム / 翻訳後修飾 / リン酸化 / ユビキチン化 / 腎臓 / 尿管 / 糸球体 / 濾過 / プロテオミクス / 免疫沈降

Outline of Annual Research Achievements

網羅的タンパク質（プロテオーム）解析でタンパク質の翻訳後修飾、特にリン酸化修飾やユビキチン化修飾の関連を研究するプロジェクトを開始した。初めに、腎臓のタンパク質をモデルに、網羅性を高めるために、腎臓のタンパク質をOff-Gel 等電点電気泳動法で分画後、トリプシン処理し、そのペプチドを質量分析装置で解析し、全分画の情報を統合して腎臓プロテオームを解析した。一方、腎臓タンパク質全体を最初にトリプシン消化し、そのペプチドをOff-Gel 等電点電気泳動法で分画し、質量分析装置で解析し、全分画の情報を統合した腎臓プロテオームを取得し、前者と比較した。その結果、前者の解析法の方が、タンパク質の同定数が多い事が示されたが、ペプチドを、Off-Gel電気泳動法で分画する方法は、ITRAQ 法や TMT法などペプチドの定量法に有用な手法と考えられた。
両分画法とも、リン酸化や酸化やユビキチン化などのタンパク質の翻訳後修飾の解析が可能で、その解析は両者を相補的に用いることで、より詳細なプロテオーム解析が得られた（Anal Chem　2015）。
また、本研究で、世界で初めて尿管のプロテオームが解析され、そのタンパク質のリストが作成された（Journal of proteomics 2015とData in briefs 201６）。その情報は半定量的情報として emPAI 情報とともに、データベース化され、今後、尿管の疾患の研究に利用されると考えられた。

Research Progress Status

27年度が最終年度であるため、記入しない。

Strategy for Future Research Activity

27年度が最終年度であるため、記入しない。

Research Products

• [Int'l Joint Research] Scripps Research Institute(米国)
• [Int'l Joint Research] Suezu Canal University(エジプト)
• [Int'l Joint Research]
• [Journal Article] Comprehensive data analysis of human ureter proteome (2016)
  • Author(s): Magdeldin S, Hirao Y, El Guoshy A, Xu B, Zhang Y, Fujinaka H, Yamamoto K, Yates JR 3rd, Yamamoto T.
  • Journal Title: Data Brief. Volume: 3 Pages: 853-857
  • DOI: 10.1016/j.dib.2016.01.050
  • Peer Reviewed / Open Access / Int'l Joint Research
• [Journal Article] A proteomic glimpse into human ureter proteome. (2016)
  • Author(s): Magdeldin S, Hirao Y, Elguoshy A, Xu B, Zhang Y, Fujinaka H, Yamamoto K, Yates JR 3rd, Yamamoto T
  • Journal Title: Proteomics Volume: 16 Issue: 1 Pages: 80-84
  • DOI: 10.1002/pmic.201500214
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Complementary protein and peptide OFFGEL fractionation for high-throughput proteomic analysis (2015)
  • Author(s): Magdeldin S, Elguoshy A, Yoshida Y, Hirao Y, Xu B, Zhang Y, Yamamoto K, Takimoto H, Fujinaka H, Kinoshita N, Yamamoto T
  • Journal Title: Anal Chem Volume: 18 Issue: 16 Pages: 8481-8488
  • DOI: 10.1021/acs.analchem.5b01911
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] .Basics and recent advances of two dimensional- polyacrylamide gel electrophoresis (2014)
  • Author(s): Magdeldin S, Enany S, Yoshida Y, Xu B, Zhang Y, Zureena Z, Lokamani I, Yaoita E, Yamamoto T
  • Journal Title: Clin Proteomics Volume: 11 Issue: 1 Pages: 16-16
  • DOI: 10.1186/1559-0275-11-16
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Deep proteome mapping of mouse kidney based on OFFGel prefractionation reveals remarkable protein post- translational modifications. (2014)
  • Author(s): Magdeldin S, Yamamoto K, Yoshida Y, Xu B, Zhang Y, Fujinaka H, Yaoita E, Yates JR 3rd, Yamamoto T
  • Journal Title: J Proteome Res. Volume: 13 Issue: 3 Pages: 1636-46
  • DOI: 10.1021/pr401122m
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Heparin increasing podocyte-specific gene expressions. (2014)
  • Author(s): Yaoita E, Yoshida Y, Nameta M, Zhang Y, Fujinaka H, Magdeldin S, Xu B, Yamamoto T.
  • Journal Title: Nephrology (Carlton) Volume: 19 Issue: 4 Pages: 195-201
  • DOI: 10.1111/nep.12207
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Decreased urinary calbindin 1 levels in proteinuric rats and humans with distal nephron segment injuries. (2014)
  • Author(s): Iida T, Fujinaka H, Xu B, Zhang Y, Magdeldin S, Nameta M, Liu Z, Yoshida Y, Yaoita E, Tomizawa S, Saito A, Yamamoto T
  • Journal Title: Clin Exp Nephrol. Volume: 18 Issue: 3 Pages: 432-43
  • DOI: 10.1007/s10157-013-0835-3
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] The State of the Human Proteome in 2013 as viewed through PeptideAtlas: Comparing the Kidney, Urine, and Plasma Proteomes for the Biology and Disease-driven Human Proteome Project. (2014)
  • Author(s): Farrah T, Deutsch EW, Omenn GS, Sun Z, Watts JD, Yamamoto T, Shteynberg D, Harris MM, Moritz RL.
  • Journal Title: J Proteome Res. Volume: 13 Issue: 02 Pages: 60-75
  • DOI: 10.4236/ojgas.2014.42011
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Presentation] Behavioral and Proteomic Analysis of Stress Response in Zebrafish (Danio rerio) (2015)
  • Author(s): Sameh Magdeldin
  • Organizer: HUPO2015
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2015-09-27
  • Int'l Joint Research
• [Presentation] Protein Post-Translational Modifications of Mouse Kidney using OFFGel Prefractionatio (2015)
  • Author(s): Sameh Magdeldin
  • Organizer: HUPO2015
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2015-09-27
  • Int'l Joint Research
• [Presentation] Urinary Proteins Originating Uniquely from Each Nephron Segment (2015)
  • Author(s): Tadashi Yamamoto
  • Organizer: HUPO2015
  • Place of Presentation: Vancouver, Canada
  • Year and Date: 2015-09-27
  • Int'l Joint Research / Invited
• [Presentation] Preparation of proteins and peptides by MS analysis in biomarker discover using human urine. (2015)
  • Author(s): Sameh Magdeldin
  • Organizer: 日本プロテオーム学会2015
  • Place of Presentation: くまもと森都心プラザ(熊本市)
  • Year and Date: 2015-07-24
• [Presentation] Application of Proteomics to Translational Nephrology (2014)
  • Author(s): Tadashi Yamamoto
  • Organizer: 1st-International Caparica Conference In Translational Nephrology & 18th Portuguese Symposium on Renal Diseases
  • Place of Presentation: Caparica, Lisbon, Portugal
  • Year and Date: 2014-10-10 – 2014-10-11
  • Invited
• [Presentation] MS- and Ab-Based Proteomics to Understand Biology and Disease of Kidney (2014)
  • Author(s): Tadashi Yamamoto, Bo Xu, Ying Zhang, Sameh Magdeldin, Masato Habuka, Hidehiko Fujinaka, Keiko Yamamoto, Yutaka Yoshida, Shin Goto, Ichiei Narita
  • Organizer: 13th HUPO World Conference
  • Place of Presentation: Madrid, Spain
  • Year and Date: 2014-10-07 – 2014-10-08
  • Invited
• [Presentation] Strategy of Discovery and Validation of Urine Biomarkers for Kidney Diseases (2014)
  • Author(s): Tadashi Yamamoto
  • Organizer: 7th AOHUPO
  • Place of Presentation: Bangkok, Thailand
  • Year and Date: 2014-08-06 – 2014-08-08
  • Invited
• [Remarks] 生体液バイオマーカーセンター
  • URL: http://bb-c.gs.niigata-u.ac.jp/about.html