Development of a reliable peptidomics technique using a novel chemical derivatization approach

• Project/Area Number: 25513001
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: オミクス計測科学
• Research Institution: Kyoto University
• Principal Investigator: Miyagawa Hisashi 京都大学, (連合)農学研究科(研究院), 教授 (10219735)
• Co-Investigator(Renkei-kenkyūsha): MIYASHITA Masahiro 京都大学, 大学院農学研究科, 助教 (80324664)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2013: ¥3,120,000 (Direct Cost: ¥2,400,000、Indirect Cost: ¥720,000)
• Keywords: ペプチド / 質量分析 / アミノ酸配列決定 / 化学修飾 / 抗菌性 / de novo sequencing / derivatization / mass spectrometry / fragmentation / peptide / ペプチドミクス / de novo シーケンス / MS/MS / 高エネルギーCID

Outline of Final Research Achievements

Peptide sequencing by mass spectrometry is known to be facilitated by chemical modification of the peptide N-terminus. In this study, we fist optimized the method for N-terminal selective modification and then investigated the modification structures suitable for Leu/Ile discrimination under high-energy CID conditions. In addition, we successfully identified three novel antimicrobial peptides from the scorpion Isometrus maculatus venom using the method developed in this study.

Research Products

• [Journal Article] Differential ¹⁴N/¹⁵N-Labeling of Peptides Using N-Terminal Charge Derivatization with a High-Proton Affinity for Straightforward <i>de novo</i> Peptide Sequencing (2013)
  • Author(s): Yoichiro Nihashi, Masahiro Miyashita, Hiroyuki Awane, and Hisashi Miyagawa
  • Journal Title: Mass Spectrometry Volume: 2 Issue: 1 Pages: A0024-A0024
  • DOI: 10.5702/massspectrometry.A0024
  • NAID: 130003388684
  • ISSN: 2186-5116, 2187-137X
  • Peer Reviewed
• [Presentation] Identification of novel antimicrobial peptides from the venom of the scorpion Isometrus maculatus (2015)
  • Author(s): Atsushi Kitanaka, Mao Yakio, Masahiro Miyashita, Yoshiaki Nakagawa and Hisashi Miyagawa
  • Organizer: 第52回ペプチド討論会
  • Place of Presentation: 平塚市中央公民館
  • Year and Date: 2015-11-16
• [Presentation] マダラサソリの毒液に含まれる抗菌性ペプチドの探索 (2015)
  • Author(s): 北中淳史・ 宮下正弘・ 中川好秋・ 宮川恒
  • Organizer: 農薬デザイン研究会
  • Place of Presentation: メルパルク京都
  • Year and Date: 2015-11-12
• [Presentation] マダラサソリの毒液に含まれる抗菌性ペプチドのde novoシーケンス解析 (2015)
  • Author(s): 北中淳史・ 宮下正弘・ 中川好秋・ 宮川恒
  • Organizer: 第63回質量分析総合討論会
  • Place of Presentation: つくば国際会議場 エポカルつくば
  • Year and Date: 2015-06-17
• [Presentation] 高エネルギーCID MS/MS分析によるペプチド中のLeu/Ileの判別におけるN末端修飾の効果 (2014)
  • Author(s): 北中淳史、 宮下正弘、 久保歩、 佐藤貴弥、豊田岐聡、 宮川恒
  • Organizer: 第62回質量分析総合討論会
  • Place of Presentation: ホテル阪急エキスポパーク
  • Year and Date: 2014-05-14
• [Presentation] MS/MS 分析によるペプチド中の Leu/Ile の判別における N 末端修飾の効果 (2014)
  • Author(s): 北中 淳史、宮下 正弘、宮川 恒
  • Organizer: 日本農芸化学会2014年度大会
  • Place of Presentation: 明治大学