Determination of the Androgenicity of Ligands Using a Single-chain Probe Carrying Androgen Receptor N-Terminal Peptides

• KIM Sung Bae

  Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST)

• UMEZAWA Yoshio

  Research Institute of Pharmaceutical Sciences, Musashino University

• TAO Hiroaki

  Research Institute for Environmental Management Technology, National Institute of Advanced Industrial Science and Technology (AIST)

抄録

The present study demonstrates a single-molecular bioluminescent probe carrying functional peptides in the N-terminal domain of the androgen receptor (AR NTD) with an improved sensorial property to androgens. The N-terminal peptides in AR were genetically fused to the ligand binding domain of AR (AR LBD) with a flexible linker, and then sandwiched between the N- and C-terminal fragments of split-firefly luciferase (FLuc) dissected at D415. We found that the proline-rich region in AR NTD efficiently interacts with AR LBD and exerts (i) an enhanced signal-to-background ratio and (ii) discrimination between agonists and antagonists with (iii) a 100-times improved sensitivity to androgens, upon comparison with previous references. A deletion mutation to the proline-rich region in AR revealed that this region is critical for the transcriptional activities. The quantum yields of these single-chain probes were estimated to be 37.8 ± 0.6%. This monomeric AR LBD-peptide binding is necessary, and sufficient for discriminating an agonist and an antagonist, where the dimerization of AR LBD is not involved. The present study guides a fundamental methodology on how to discriminate weak protein-peptide binding, and provides a new insight into the contribution of functional peptides in AR NTD to the initial activation of monomeric AR.

収録刊行物

• Analytical Sciences

  Analytical Sciences 25 (12), 1415-1420, 2009

  社団法人 日本分析化学会

キーワード

• Analytical Chemistry

詳細情報

• CRID: 1390282679232376576
• NII論文ID: 10026780403
• NII書誌ID: AA10500785
• DOI: 10.2116/analsci.25.1415
• COI: 1:STN:280:DC%2BD1MfisFaktw%3D%3D
• ISSN: 13482246
• ISSN: 09106340
• NDL書誌ID: 10465439
• PubMed: 20009327
• Web Site: https://ndlsearch.ndl.go.jp/books/R000000004-I10465439
• Web Site: https://link.springer.com/content/pdf/10.2116/analsci.25.1415.pdf
• Web Site: https://link.springer.com/article/10.2116/analsci.25.1415/fulltext.html
• Web Site: http://www.jstage.jst.go.jp/article/analsci/25/12/25_12_1415/_pdf
• 本文言語コード: en
• データソース種別:
  • JaLC
  • NDL
  • Crossref
  • PubMed
  • CiNii Articles
• 抄録ライセンスフラグ: 使用不可