Development of a method to screen MS-silent post-translational modifications

• Project/Area Number: 24590044
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Physical pharmacy
• Research Institution: Tohoku University
• Principal Investigator: GOTO Takaaki 東北大学, 薬学研究科(研究院), 講師 (40344684)
• Co-Investigator(Kenkyū-buntansha): OE Tomoyuki 東北大学, 大学院薬学研究科, 教授 (10203712) ; LEE Seon Hwa 東北大学, 大学院薬学研究科, 助教 (60519776)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Project Status: Completed (Fiscal Year 2014)
• Budget Amount: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2014: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2013: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000); Fiscal Year 2012: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: 分析科学 / 生体分子 / タンパク質 / 質量分析 / 翻訳後修飾 / プロテオーム

Outline of Final Research Achievements

A novel methodology to screen MS-silent post-translational modifications on protein has been examined. This method is based on MS-based proteomics combined with the group-specific immunoaffinity extraction, complementary data analyses in both positive and negative mode, and LC/high-resolution MS. This strategy can be used for screening of modifications including isomerization.

Research Products

• [Journal Article] Mass spectrometric characterizations of protein carbonylation: Comparison between three different conditions, oxidation by CuII/ascorbic acid, adduction of methyl glyoxal, and adduction of 4-hydroxy-2(E)-nonenal (2014)
  • Author(s): Kazuki Maeda, Hiroyuki Funatsu, Yuki Miyazawa, Yasushi Misawa, Takaaki Goto, Seon Hwa Lee and Tomoyuki Oe
  • Journal Title: International Journal of Mass Spectrometry Volume: 373 Pages: 72-80
  • DOI: 10.1016/j.ijms.2014.09.012
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Predicted multiple selected reaction monitoring to screen activated drug-mediated modifications on human serum albumin. (2014)
  • Author(s): Fumio Osaki, Takaaki Goto, Seon Hwa Lee and Tomoyuki Oe
  • Journal Title: Analytical Biochemistry Volume: 449 Pages: 59-67
  • DOI: 10.1016/j.ab.2013.12.016
  • Peer Reviewed
• [Journal Article] Complete amino acid sequencing and immunoaffinity clean-up can facilitate screening of various chemical modifications on human serum albumin. (2013)
  • Author(s): Takaaki Goto, Kazuyuki Murata, Seon Hwa Lee and Tomoyuki Oe
  • Journal Title: Analytical and Bioanalytical Chemistry Volume: 405 Issue: 23 Pages: 7383-7395
  • DOI: 10.1007/s00216-013-7146-0
  • Peer Reviewed
• [Journal Article] Aldehyde stress-mediated novel modification of proteins: epimerization of the N-terminal amino acid. (2013)
  • Author(s): Ryo Kajita, Takaaki Goto, Seon Hwa Lee and Tomoyuki Oe
  • Journal Title: Chemical Research in Toxicology Volume: 26 Issue: 12 Pages: 1926-1936
  • DOI: 10.1021/tx400354d
  • Peer Reviewed
• [Journal Article] Can Edman degradation be used for quantification? Isotope-dilution liquid chromatography-electrospray ionization tandem mass spectrometry and the long-term stability of 20 phenylthiohydantoin-amino acids (2013)
  • Author(s): Satoh, R., Goto, T., Lee, S.H., Oe, T.
  • Journal Title: Anal. Bioanal. Chem Volume: (in press) Issue: 25 Pages: 8001-8010
  • DOI: 10.1007/s00216-013-6901-6
  • Peer Reviewed
• [Presentation] 血清アルブミン上の網羅的化学修飾解析を指向したLC/MSによる手法の開発 (2014)
  • Author(s): 工藤裕太、李宣和、後藤貴章、大江知行
  • Organizer: 第53回日本薬学会東北支部大会
  • Place of Presentation: いわき
  • Year and Date: 2014-10-05
• [Presentation] 血清アルブミンの化学修飾オミクス研究：新規バイオマーカー開発に向けて (2014)
  • Author(s): 後藤貴章
  • Organizer: 平成２６年度日本薬学会東北支部 第3回 物理・分析系若手研究者セミナー
  • Place of Presentation: 仙台
  • Year and Date: 2014-09-20
  • Invited
• [Presentation] Simultaneous mass spectrometric analysis of various chemical modifications on human serum albumin: strategies for clean-up, sequence coverage, and identification (2014)
  • Author(s): Takaaki Goto, Yuta Kudo, Kazuyuki Murata, Seon Hwa Lee and Tomoyuki Oe
  • Organizer: 62nd ASMS Conference on Mass Spectrometry and Allied Topics
  • Place of Presentation: Baltimore, USA
  • Year and Date: 2014-06-15 – 2014-06-19
• [Presentation] 血清アルブミン上の網羅的化学修飾解析を目的とした高分解能LC/MSによる修飾ペプチド検出法の検討 (2014)
  • Author(s): 後藤貴章、工藤裕太、李宣和、大江知行
  • Organizer: 第74回分析化学討論会
  • Place of Presentation: 郡山
  • Year and Date: 2014-05-24 – 2014-05-25
• [Presentation] シッフ塩基形成を介した血清アルブミンN末端アミノ酸の光学反転 (2013)
  • Author(s): 梶田遼、後藤貴章、李宣和、大江知行
  • Organizer: 平成25年度日本分析化学会東北支部若手交流会
  • Place of Presentation: 仙台
• [Presentation] 標品不要のタンパク質・ペプチド絶対定量法の開発: エドマン分解反応物の同位体希釈LC-ESI/MS/MS法 (2013)
  • Author(s): 佐藤涼、後藤貴章、李宣和、大江知行
  • Organizer: 平成25年度日本分析化学会東北支部若手交流会
  • Place of Presentation: 仙台
• [Presentation] アルデヒドによるN末端アミノ酸の異性化における反応性の検討 (2013)
  • Author(s): 東由佳、後藤貴章、李宣和、大江知行
  • Organizer: 第52回日本薬学会東北支部大会
  • Place of Presentation: 仙台
• [Presentation] Predicted Selected Reaction Monitoring を用いたヘモグロビン上の化学修飾スクリーニング法の開発 (2013)
  • Author(s): 小島弘貴、李宣和、後藤貴章、大江知行
  • Organizer: 第52回日本薬学会東北支部大会
  • Place of Presentation: 仙台
• [Presentation] アルデヒドによるタンパク質N末端アミノ酸異性化の研究 (2013)
  • Author(s): 梶田遼、李宣和、後藤貴章、大江知行
  • Organizer: 新アミノ酸分析研究会 第3回学術講演会
  • Place of Presentation: 東京
• [Presentation] Mechanistic Study on Non-enzymatic Isomerization of N-terminal Amino Acids by Endogenous Aldehydes (2012)
  • Author(s): Ryo Kajita, Takaaki Goto, Seon Hwa Lee and Tomoyuki Oe
  • Organizer: 19th International Mass Spectrometry Conferrence
  • Place of Presentation: Kyoto, Japan
• [Presentation] Isotope dilution LC-ESI/MS/MS method for the determination of phenylthiohydantoin (PTH)-amino acids derived from Edman degradation products of proteins (2012)
  • Author(s): Ryo Satoh, Takaaki Goto, Seon Hwa Lee and Tomoyuki Oe
  • Organizer: 19th International Mass Spectrometry Conferrence
  • Place of Presentation: Kyoto, Japan
• [Presentation] Chemical modificomics ～Alternative analytical platform for efficient biomarker discovery～ (2012)
  • Author(s): Tomoyuki Oe, Takaaki Goto, Shota Kojima, Shohei Shitamichi, Kazuyuki Murata, Kohei Miyamoto and Seon Hwa Lee
  • Organizer: 19th International Mass Spectrometry Conferrence
  • Place of Presentation: Kyoto, Japan
• [Presentation] N末端アミノ酸のpyridoxal 5'-phosphateに対する反応性と非酵素的異性化 (2012)
  • Author(s): 梶田遼、東由佳、後藤貴章、李宣和、大江知行
  • Organizer: 第25回バイオメディカル分析科学シンポジウム
  • Place of Presentation: 東京
• [Presentation] タンパク質のエドマン分解反応物: PTH-アミノ酸を用いた同位体希釈LC/MS法の構築 (2012)
  • Author(s): 佐藤涼、後藤貴章、李宣和、大江知行
  • Organizer: 新アミノ酸分析研究会 第二回学術講演会
  • Place of Presentation: 東京