Open AccessConsideration on Efficient Recombinant Protein Production: Focus on Substrate Protein-Specific Compatibility Patterns of Molecular Chaperones
Author(s) -
Naohiro Yano,
T. Emi,
David Gregory,
Alexey V. Fedulov
Publication year - 2021
Publication title -
the protein journal
Language(s) - English
Resource type - Journals
eISSN - 1875-8355
pISSN - 1572-3887
DOI - 10.1007/s10930-021-09995-4
Subject(s) - groel , chaperone (clinical) , groes , protein folding , chaperonin , recombinant dna , foldase , biochemistry , chemical chaperone , escherichia coli , biology , microbiology and biotechnology , chemistry , gene , unfolded protein response , medicine , pathology
Expression of recombinant proteins requires at times the aid of molecular chaperones for efficient post-translational folding into functional structure. However, predicting the compatibility of a protein substrate with the right type of chaperone to produce functional proteins is a daunting issue. To study the difference in effects of chaperones on His-tagged recombinant proteins with different characteristics, we performed in vitro proteins expression using Escherichia coli overexpressed with several chaperone 'teams': Trigger Factor (TF), GroEL/GroES and DnaK/DnaJ/GrpE, alone or in combinations, with the aim to determine whether protein secondary structure can serve as predictor for chaperone success. Protein A, which has a helix dominant structure, showed the most efficient folding with GroES/EL or TF chaperones alone, whereas Protein B, which has less helix in the structure, showed a remarkable effect on the DnaK/J/GrpE system alone. This tendency was also seen with other recombinant proteins with particular properties. With the chaperons' assistance, both proteins were synthesized more efficiently in the culture at 22.5 °C for 20 h than at 37 °C for 3 h. These findings suggest a novel avenue to study compatibility of chaperones with substrate proteins and optimal culture conditions for producing functional proteins with a potential for predictive analysis of the success of chaperones based on the properties of the substrate protein.