• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Visualization of molecular interactions at the nanometer scale

Research Project

Project/Area Number 25650062
Research Category

Grant-in-Aid for Challenging Exploratory Research

Allocation TypeMulti-year Fund
Research Field Cell biology
Research InstitutionNagoya University

Principal Investigator

FUJIMOTO Toyoshi  名古屋大学, 医学(系)研究科(研究院), 教授 (50115929)

Project Period (FY) 2013-04-01 – 2015-03-31
Project Status Completed (Fiscal Year 2014)
Budget Amount *help
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Fiscal Year 2013: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
Keywords生体膜 / 電子顕微鏡 / 膜タンパク質 / 凍結割断レプリカ / 膜蛋白質 / 凍結割断レプリカ法 / ビオチン化
Outline of Final Research Achievements

Methods to visualize the location where two different proteins interact include fluorescence resonance energy transfer, bimolecular fluorescence complementation, and proximity ligation assay, but their spatial resolution is not high. In this study, we conceived an idea of a method based on quick-freezing and freeze-fracture replica labeling method. The principle of the method was: 1) tagging of biotinylation enzyme BirA and its acceptor peptide AcP to the cytoplasmic domain of two transmembrane proteins A and B, respectively; 2) biotinylation of AcP in freeze-fracture replicas; 3) immunolabeling and electron microscopic observation of biotins. Although various conditions were examined, satisfactory results were not obtained so far. Further trials using different conditions are needed.

Report

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

    (2 results)

All 2014 Other

All Journal Article (1 results) (of which Peer Reviewed: 1 results) Remarks (1 results)

  • [Journal Article] Microscopic methods to observe the distribution of lipids in the cellular membrane2014

    • Author(s)
      Sho Takatori, Rob Mesman, Toyoshi Fujimoto
    • Journal Title

      Biochemistry

      Volume: Vol.53(4) Issue: 4 Pages: 639-653

    • DOI

      10.1021/bi401598v

    • Related Report
      2014 Annual Research Report
    • Peer Reviewed
  • [Remarks] 名古屋大学大学院医学系研究科・分子細胞学分野ホームページ

    • URL

      http://www.med.nagoya-u.ac.jp/cel-bio/index-j.html

    • Related Report
      2014 Annual Research Report

URL: 

Published: 2014-07-25   Modified: 2019-07-29  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi