Intramembrane proteolysis of beta-APP in a gamma-secretase independent mechanism

• Project/Area Number: 26461745
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Psychiatric science
• Research Institution: Osaka University
• Principal Investigator: Yanagida Kanta 大阪大学, 医学系研究科, 特任研究員(常勤) (70467596)
• Co-Investigator(Kenkyū-buntansha): 田上 真次 大阪大学, 医学系研究科, 助教 (40362735) ; 大河内 正康 大阪大学, 医学系研究科, 講師 (90335357)
• Project Period (FY): 2014-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2016: ¥1,300,000 (Direct Cost: ¥1,000,000、Indirect Cost: ¥300,000); Fiscal Year 2015: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2014: ¥1,950,000 (Direct Cost: ¥1,500,000、Indirect Cost: ¥450,000)
• Keywords: アルツハイマー病 / γ-セクレターゼ / CRISPR/CAS9 / アミロイド / γ‐セクレターゼ

Outline of Final Research Achievements

To investigate the γ-secretase independent intramembrane cleavage mechanism of amyloid β precursor protein (βAPP), we generated presenilin 1 and presenilin 2 double knockout (PSDKO) cells by CRISPR/Cas9 system. PSDKO cells were treated with various protease inhibitors and intracellular oligopeptides derived from transmembrane domain of βAPP were quantified by LC-Ms/Ms. As a result, eight tripeptides were decreased by treatment of lysosomal enzyme inhibitor chloroquine. Since these peptides were not decreased by inhibition of cathepsins, the major lysosomal proteases, it was suggested that intramembrane domain of βAPP was degraded by tripeptidyl peptidase 1, which releases tripeptides from N-termini of proteins.

Research Products

• [Journal Article] Semagacestat Is a Pseudo-Inhibitor of γ-Secretase (2017)
  • Author(s): Tagami S, Yanagida K, Kodama TS, Takami M, Mizuta N, Oyama H, Nishitomi K, Chiu YW, Okamoto T, Ikeuchi T, Sakaguchi G, Kudo T, Matsuura Y, Fukumori A, Takeda M, Ihara Y, Okochi M.
  • Journal Title: Cell Rep Volume: 21 Issue: 1 Pages: 259-273
  • DOI: 10.1016/j.celrep.2017.09.032
  • Peer Reviewed / Open Access
• [Presentation] CRISPR/CAS9法によるPresenilin1/Presenilin2ダブルノックアウト細胞の作製 (2015)
  • Author(s): 柳田寛太
  • Organizer: 第34回日本認知症学会学術集会
  • Place of Presentation: リンクステーションホール青森
  • Year and Date: 2015-10-03
• [Presentation] γセクレターゼの阻害と促進 (2014)
  • Author(s): 大河内正康
  • Organizer: 第33回日本認知症学会学術集会
  • Place of Presentation: パシフィコ横浜
  • Year and Date: 2014-11-30