Studies on construction of a novel cytotoxic functional molecule using pore-forming lectin

2018 Fiscal Year Final Research Report

• Project/Area Number: 16K07685
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Ibaraki University
• Principal Investigator: KOUZUMA Yoshiaki 茨城大学, 農学部, 教授 (10284556)
• Co-Investigator(Kenkyū-buntansha): 中島 崇 九州大学, 農学研究院, 助教 (20380553)
• Project Period (FY): 2016-04-01 – 2019-03-31
• Keywords: 小孔形成レクチン / ガン細胞傷害

Outline of Final Research Achievements

In the present study, we tried to construct a novel cytotoxic functional molecule with controlling the activities using pore-forming lectin, CEL-III. Among six CEL-III mutants in which two cysteine residues were introduced for activity control, only E331C/S351C mutant did not exert the hemolytic activity in the absence of a reducing agent and the activity was recovered in the presence of it. Furthermore, E331C/S351C mutant showed proliferation inhibition activities toward K562 and MOLT-4 cells only in the presence of reducing agent (GSH) significantly. On the other hand, we were able to obtain CRD1 mutans of CEL-III that might bind to cancer cells specifically using phage-display method.

Free Research Field

生物化学

Academic Significance and Societal Importance of the Research Achievements

本研究により、小孔形成レクチンCEL-IIIのE331とS350にCys残基を導入することで細胞等に対する活性を還元剤によって制御することが可能になり、ファージディスプレイ法を用いて得られたガン化細胞特異的に結合するCRD変異体の糖結合部位の配列を上記のCEL-IIIに組み込むことによって、還元剤による活性制御が可能な新規ガン化細胞傷害機能性分子の構築が可能になり、ガン細胞除去などのためのツールとしての応用が期待できる。