THETA system allows one-step isolation of tagged proteins through temperature-dependent protein–peptide interaction

Kota Miura, Yusuke Tsuji, Hiromasa Mitsui, Takuya Oshima, Yosei Noshi, Yudai Arisawa, Keiko Okano, Toshiyuki Okano

Research output: Contribution to journalArticle

Abstract

Tools to control protein-protein interactions by external stimuli have been extensively developed. For this purpose, thermal stimulation can be utilized in addition to light. In this study, we identify a monoclonal antibody termed C13 mAb, which shows an approximately 480-fold decrease in the affinity constant at 37 °C compared to that at 4 °C. Next, we apply this temperature-dependent protein-peptide interaction for one-step protein purifications. We term this THermal-Elution-based TAg system as the THETA system, in which gel-immobilized C13 mAb-derived single-chain variable fragment (scFv) (termed THETAL) is able to bind with proteins tagged by C13 mAb-epitope(s) (THETAS) at 4 °C and thermally release at 37–42 °C. Moreover, to reveal the temperature-dependent interaction mechanism, molecular dynamics simulations are performed along with epitope mapping experiments. Overall, the high specificity and reversibility of the temperature-dependent features of the THETA system will support a wide variety of future applications such as thermogenetics.

Original languageEnglish
Article number207
JournalCommunications Biology
Volume2
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Temperature
heat
temperature
Proteins
proteins
molecular dynamics
protein-protein interactions
Epitopes
Hot Temperature
epitopes
monoclonal antibodies
Epitope Mapping
Single-Chain Antibodies
gels
peptides
Molecular Dynamics Simulation
Purification
Molecular dynamics
Gels
Monoclonal Antibodies

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Medicine (miscellaneous)

Cite this

THETA system allows one-step isolation of tagged proteins through temperature-dependent protein–peptide interaction. / Miura, Kota; Tsuji, Yusuke; Mitsui, Hiromasa; Oshima, Takuya; Noshi, Yosei; Arisawa, Yudai; Okano, Keiko; Okano, Toshiyuki.

In: Communications Biology, Vol. 2, No. 1, 207, 01.12.2019.

Research output: Contribution to journalArticle

Miura, Kota ; Tsuji, Yusuke ; Mitsui, Hiromasa ; Oshima, Takuya ; Noshi, Yosei ; Arisawa, Yudai ; Okano, Keiko ; Okano, Toshiyuki. / THETA system allows one-step isolation of tagged proteins through temperature-dependent protein–peptide interaction. In: Communications Biology. 2019 ; Vol. 2, No. 1.
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