Domain-wise dissection of thermal stability enhancement in multidomain proteins

• Published in: International journal of biological macromolecules Vol. 237; p. 124141
• Main Authors: Oh, Jisung, Durai, Prasannavenkatesh, Kannan, Priyadharshini, Park, Jaehui, Yeon, Young Joo, Lee, Won-Kyu, Park, Keunwan, Seo, Moon-Hyeong
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.05.2023
• Subjects: Enzyme Stability; Glutamine-binding protein; Hydroxyacyl-coenzyme A dehydrogenase; Melting temperature; Mutant Proteins - chemistry; Protein engineering; Protein thermal stability; Proteins - chemistry; Tyrosine-protein kinase SYK; Tyrosine-protein kinase SYK; Protein engineering; Protein thermal stability; Melting temperature; Glutamine-binding protein; Hydroxyacyl-coenzyme A dehydrogenase
• ISSN: 0141-8130; 1879-0003
• DOI: 10.1016/j.ijbiomac.2023.124141

Stability is critical for the proper functioning of all proteins. Optimization of protein thermostability is a key step in the development of industrial enzymes and biologics. Herein, we demonstrate that multidomain proteins can be stabilized significantly using domain-based engineering followed by the recombination of the optimized domains. Domain-level analysis of designed protein variants with similar structures but different thermal profiles showed that the independent enhancement of the thermostability of a constituent domain improves the overall stability of the whole multidomain protein. The crystal structure and AlphaFold-predicted model of the designed proteins via domain-recombination provided a molecular explanation for domain-based stepwise stabilization. Our study suggests that domain-based modular engineering can minimize the sequence space for calculations in computational design and experimental errors, thereby offering useful guidance for multidomain protein engineering. •Thermostability of multidomain proteins was analyzed on a domain-by-domain basis.•Stability of a constituent domain determines overall stability of the protein.•Domain-based approach helps stabilize multidomain proteins.