Ligand-directed acyl imidazole chemistry for labeling of membrane-bound proteins on live cells

Sho Hei Fujishima, Ryosuke Yasui, Takayuki Miki, Akio Ojida, Itaru Hamachi

研究成果: ジャーナルへの寄稿記事

79 引用 (Scopus)

抄録

Chemistry-based protein labeling in living cells is undoubtedly useful for understanding natural protein functions and for biological/pharmaceutical applications. Here, we report a novel approach for endogenous membrane-bound protein labeling for both in vitro and live cell conditions. A moderately reactive alkyloxyacyl imidazole (AI) assisted by ligand-binding affinity (ligand-directed AI (LDAI)) chemistry allowed us to selectively modify natural proteins, such as dihydrofolate reductase (DHFR) and folate receptor (FR), neither of which could be efficiently labeled using the recently developed ligand-directed tosylate approach. It was clear that LDAI selectively labeled a single Lys(K32) in DHFR, proximal to the ligand-binding pocket. We also demonstrate that the fluorescein-labeled (endogenous, by LDAI) FR works as a fluorescent biosensor on the live KB cell surface, which allowed us to carry out unprecedented in situ kinetic analysis of ligand binding to FR.

元の言語英語
ページ(範囲)3961-3964
ページ数4
ジャーナルJournal of the American Chemical Society
134
発行部数9
DOI
出版物ステータス出版済み - 3 7 2012

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Labeling
Membrane Proteins
Ligands
Proteins
Membranes
Folic Acid
Tetrahydrofolate Dehydrogenase
KB Cells
Biosensing Techniques
imidazole
Fluorescein
Biosensors
Drug products
Cells
Kinetics
Pharmaceutical Preparations

All Science Journal Classification (ASJC) codes

  • Catalysis
  • Chemistry(all)
  • Biochemistry
  • Colloid and Surface Chemistry

これを引用

Ligand-directed acyl imidazole chemistry for labeling of membrane-bound proteins on live cells. / Fujishima, Sho Hei; Yasui, Ryosuke; Miki, Takayuki; Ojida, Akio; Hamachi, Itaru.

:: Journal of the American Chemical Society, 巻 134, 番号 9, 07.03.2012, p. 3961-3964.

研究成果: ジャーナルへの寄稿記事

Fujishima, Sho Hei ; Yasui, Ryosuke ; Miki, Takayuki ; Ojida, Akio ; Hamachi, Itaru. / Ligand-directed acyl imidazole chemistry for labeling of membrane-bound proteins on live cells. :: Journal of the American Chemical Society. 2012 ; 巻 134, 番号 9. pp. 3961-3964.
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