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study of low-immunogenic monomeric streptavidin

Research Project

Project/Area Number 25640088
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Tumor therapeutics
Research InstitutionThe University of Tokyo

Principal Investigator

DOI Hirofumi  東京大学, 先端科学技術研究センター, 特任教授 (80403335)

Co-Investigator(Kenkyū-buntansha) SUGIYAMA Akira  東京大学, アイソトープ総合センター, 助教 (40562715)
Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000)
Fiscal Year 2014: ¥650,000 (Direct Cost: ¥500,000、Indirect Cost: ¥150,000)
Fiscal Year 2013: ¥3,380,000 (Direct Cost: ¥2,600,000、Indirect Cost: ¥780,000)
Keywordsテトラマー / モノマー / 巻き戻し / ビオチン結合 / 分子動力学 / シミュレ=ション / 塩橋 / シミュレーション / ストレプトアビジン / 低免疫原性 / モノマー化
Outline of Final Research Achievements

Based on the low immunogenicity streptavidin LISA314 we tried to make a monomer of LISA314. Hydrophobic amino acids in the tetramer interfaces, or in the dimer interfaces were replaced by Thr or Ser soluble amino acids. Also, 120Trp was replaced to Gly. Genes introducing these mutations (with His tag) was artificially synthesized, and then LISA314 monomers were expressed in E. coli and purified. The binding affinity of the monomer with biotin was measured using a Biacore system, but binding affinity was lost.
In order to explore the cause, we carried out molecular dynamics simulations in the monomer state. The simulation suggested that the interaction between two amino acids that form a salt bridge is important for biotin binding.

Report

(3 results)
  • 2014 Annual Research Report   Final Research Report ( PDF )
  • 2013 Research-status Report

URL: 

Published: 2014-07-25   Modified: 2021-04-07  

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