Development of a chemical screening system using aqueorin-fused protein

Masafumi Suzuki, Shoichiro Furukawa, Chikanori Kuramori, Chika Sawa, Yasuaki Kabe, Mitsuhiro Nakamura, Jun ichi Sawada, Yuki Yamaguchi, Satoshi Sakamoto, Satoshi Inouye, Hiroshi Handa

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

We developed a unique screening system that consists of combination of high photo-sensitivity of photoprotein aequorin (AQ) and our developed high-performance affinity purification system. In the present study, we demonstrated to detect the specific interaction between methotrexate (MTX) and its target dihydrofolate reductase (DHFR) fused with AQ. We succeeded to prepare highly purified AQ-fused DHFR, which showed high sensitive light emission. To test the screening system, we prepared the complex of MTX-immobilized magnetic nanobeads and AQ-fused DHFR. Bound AQ-fused DHFR with the beads was specifically released by addition of MTX. Thus, this methodology enables us to search a novel chemical that binds to target proteins without complicated processes. Furthermore, thank to the highly sensitive luminescence intensity of AQ, this methodology would be performed in very small scale with high responsibility, leading to development of high throughput screening systems.

Original languageEnglish
Pages (from-to)600-605
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume368
Issue number3
DOIs
Publication statusPublished - 2008 Apr 11
Externally publishedYes

Fingerprint

Aequorin
Tetrahydrofolate Dehydrogenase
Screening
Methotrexate
Proteins
Luminescent Proteins
Photosensitivity
Light emission
Luminescence
Purification
Throughput
Light

Keywords

  • Aequorin (AQ)
  • Affinity purification
  • Chemical screening system
  • Fusion protein
  • Magnetic nanobeads (FG beads)

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Development of a chemical screening system using aqueorin-fused protein. / Suzuki, Masafumi; Furukawa, Shoichiro; Kuramori, Chikanori; Sawa, Chika; Kabe, Yasuaki; Nakamura, Mitsuhiro; Sawada, Jun ichi; Yamaguchi, Yuki; Sakamoto, Satoshi; Inouye, Satoshi; Handa, Hiroshi.

In: Biochemical and Biophysical Research Communications, Vol. 368, No. 3, 11.04.2008, p. 600-605.

Research output: Contribution to journalArticle

Suzuki, M, Furukawa, S, Kuramori, C, Sawa, C, Kabe, Y, Nakamura, M, Sawada, JI, Yamaguchi, Y, Sakamoto, S, Inouye, S & Handa, H 2008, 'Development of a chemical screening system using aqueorin-fused protein', Biochemical and Biophysical Research Communications, vol. 368, no. 3, pp. 600-605. https://doi.org/10.1016/j.bbrc.2008.01.107
Suzuki, Masafumi ; Furukawa, Shoichiro ; Kuramori, Chikanori ; Sawa, Chika ; Kabe, Yasuaki ; Nakamura, Mitsuhiro ; Sawada, Jun ichi ; Yamaguchi, Yuki ; Sakamoto, Satoshi ; Inouye, Satoshi ; Handa, Hiroshi. / Development of a chemical screening system using aqueorin-fused protein. In: Biochemical and Biophysical Research Communications. 2008 ; Vol. 368, No. 3. pp. 600-605.
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