Generation of non-standard macrocyclic peptides specifically binding TSC-22 homologous gene-1

Sophie T PhuongDung Tran; Christopher J Hipolito; Hiroyuki Suzuki; Rudy Xie; Huynh Dam Kim Tuyen; Peter Ten Dijke; Naohiro Terasaka; Yuki Goto; Hiroaki Suga; Mitsuyasu Kato

doi:10.1016/j.bbrc.2019.06.035

Generation of non-standard macrocyclic peptides specifically binding TSC-22 homologous gene-1

Biochem Biophys Res Commun. 2019 Aug 20;516(2):445-450. doi: 10.1016/j.bbrc.2019.06.035. Epub 2019 Jun 18.

Authors

Affiliations

¹ PhD Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, 305-8575, Japan; Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, Japan.
² Department of Cancer Signaling, Faculty of Medicine, University of Tsukuba, Japan; Transborder Medical Research Center, Faculty of Medicine, University of Tsukuba, Japan.
³ Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, Japan.
⁴ Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, Japan; Graduate School of Comprehensive Human Sciences, University of Tsukuba, Japan.
⁵ Department of Cancer Signaling, Faculty of Medicine, University of Tsukuba, Japan; Department of Molecular Cell and Chemical Biology, Oncode Institute, Leiden University Medical Center, Einthovenweg 20, 2333, ZC Leiden, the Netherlands.
⁶ Department of Chemistry, Graduate School of Science, The University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033, Japan.
⁷ Department of Experimental Pathology, Faculty of Medicine, University of Tsukuba, Japan; Transborder Medical Research Center, Faculty of Medicine, University of Tsukuba, Japan. Electronic address: mit-kato@md.tsukuba.ac.jp.

PMID: 31227214
DOI: 10.1016/j.bbrc.2019.06.035

Abstract

Transforming growth factor-β 1 (TGFβ1)-stimulated clone 22 (TSC22) family includes proteins containing a leucine zipper domain and a TSC-box that are highly conserved during evolution. Currently, limited data are available on the function of this protein family, especially of TSC-22 homologous gene-1 (THG-1)/TSC22 domain family member 4 (TSC22D4). Similar to other family members, THG-1 functions depending on its interaction with the partner proteins and it is suggested to mediate a broad range of biological processes. THG-1-specific binding molecules will be instrumental for elucidating its functions. Therefore, the Random non-standard Peptide Integrated Discovery (RaPID) system was modified using commercially available materials and used for selecting macrocyclic peptides (MCPs) that bind to THG-1. Several MCPs were identified to bind THG-1. Fluorescein- and biotin-tagged MCPs were synthesized and employed as THG-1 detection probes. Notably, a fluorescein-tagged MCP specifically detected THG-1-expressing cells. Biotin-tagged MCPs can be successfully used for Enzyme-Linked Protein Sorbent Assay (ELISA) like assay of THG-1 protein and affinity-precipitation of purified THG-1 and endogenous THG-1 in esophageal squamous cell carcinoma cell lysates. The modified RaPID system rapidly and successfully identified THG-1-binding MCPs in vitro and the synthesized THG-1 binding MCPs are useful alternatives acting for antibodies.

Keywords: Affinity precipitation; ELISA-Like peptide sorbent assay; Fluorescein imaging; Non-standard macrocyclic peptide; TSC-22 homologous gene-1.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Fluorescein / metabolism
HEK293 Cells
Humans
Macrocyclic Compounds / metabolism*
Peptides / metabolism*
Transcription Factors / metabolism*

Substances

Macrocyclic Compounds
Peptides
Transcription Factors
Fluorescein