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

Autor:	Takuya Oshima, Hiromasa Mitsui, Yudai Arisawa, Yosei Noshi, Keiko Okano, Kota Miura, Toshiyuki Okano, Yusuke Tsuji
Jazyk:	angličtina
Rok vydání:	2019
Předmět:	Biomaterials - proteins medicine.drug_class Antibody Affinity Medicine (miscellaneous) Biocompatible Materials Peptide One-Step Molecular Dynamics Simulation Monoclonal antibody Article General Biochemistry Genetics and Molecular Biology Epitopes Mice Molecular dynamics Thermal stimulation Antibody Specificity Protein purification medicine Animals Biotinylation lcsh:QH301-705.5 Antibody isolation and purification chemistry.chemical_classification Mice Inbred BALB C Hybridomas Chemistry Temperature Antibodies Monoclonal Proteins Reproducibility of Results Hydrogen-Ion Concentration Epitope mapping lcsh:Biology (General) Mutation Biophysics Peptides General Agricultural and Biological Sciences Tag system Epitope Mapping
Zdroj:	Communications Biology, Vol 2, Iss 1, Pp 1-10 (2019) Communications Biology
ISSN:	2399-3642
Popis:	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. Kota Miura et al. develop the THermal-Elution-based TAg (THETA) system that allows one-step isolation of tagged proteins. The temperature-dependent protein-peptide interaction used in the THETA system ensures high specificity for protein purification.
Databáze:	OpenAIRE
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