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


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.

SUBMITTER: Miura K 

PROVIDER: S-EPMC6572768 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature

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

Miura Kota K   Tsuji Yusuke Y   Mitsui Hiromasa H   Oshima Takuya T   Noshi Yosei Y   Arisawa Yudai Y   Okano Keiko K   Okano Toshiyuki T  

Communications biology 20190614


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 a  ...[more]

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