Generation of Active Protease Depending on Peptide-Protein Interactions Using Interaction-Dependent Native Chemical Ligation and Protein Trans-Splicing

• Tsuyoshi Takahashi

  Chemistry and Chemical Biology, Graduate School of Science and Technology, Gunma University, 1-5-1 Tenjincho, Kiryu, Gunma 376-8515 , Japan

Abstract

An artificial signal transduction system has been constructed by employing engineered human immunodeficiency type-1 (HIV-1) protease and Nostoc punctiforme PCC73102 (Npu) DnaE intein. While the truncation of four amino acid residues at the N-terminus of HIV-1 protease diminished its activity, the attachment of the PQIT sequence into the truncated protease by protein trans-splicing (PTS) reconstituted the enzymatic activity. By combining interaction-dependent native chemical ligation (IDNCL) with the PTS reaction, the peptide-protein interaction was clearly detected by measuring HIV-1 protease activity. Src homology domain 2 (SH2) of c-Src (SrcSH2) and phosphopeptides were used as model binding pairs. HIV-1 protease activities were dose-dependently increased after the IDNCL-PTS reaction when the peptides containing pYEEI (pY = phosohotyrosine) and pYEE sequences were used as the input peptides. HIV-1 protease activity generated by IDNCL-PTS might activate several enzymes, and therefore, the artificial signal transduction system might be available in synthetic biology.

Journal

• Bulletin of the Chemical Society of Japan

  Bulletin of the Chemical Society of Japan 92 (10), 1767-1772, 2019-07-09

  Oxford University Press (OUP)

Keywords

• General Chemistry

Details

• CRID: 1360283694073851392
• NII Article ID: 130007728636
• DOI: 10.1246/bcsj.20190159
• ISSN: 13480634
• ISSN: 00092673
• Web Site: https://academic.oup.com/bcsj/article-pdf/92/10/1767/56299553/bcsj.20190159.pdf
• Data Source:
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