Thermostable phenylacetic acid degradation protein TtPaaI from Thermus thermophilus as a scaffold for tetravalent display of proteins

• Published in: Protein expression and purification 2025-03, Vol.227, p.106623, Article 106623
• Main Authors: Chorążewska, Aleksandra, Regan, Darragh, Kalka, Marta, Ciura, Krzysztof, Porębska, Natalia, Opaliński, Łukasz
• Format: Article
• Language: English
• Subjects: Galectin-7; Multivalent display; Oligomerization scaffold; TtPaaI
• ISSN: 1046-5928; 1096-0279; 1096-0279
• DOI: 10.1016/j.pep.2024.106623

Numerous proteins in nature strictly require oligomerization for their full activity. Moreover, the function of natural and artificial proteins can me adjusted by altering their oligomeric state, leading to development of biotechnologically-relevant biomacromolecules. Oligomerization scaffolds from natural sources and designed de novo enable shuffling the oligomeric state and valency of biomacromolecules. In this report we probed the scaffolding potential of the thermostable phenylacetic acid degradation protein acyl-CoA from Thermus thermophilus (TtPaaI). We designed and successfully produced the fusion protein between TtPaaI (scaffold) and galectin-7, a multifunctional lectin implicated in human diseases (ligand) and demonstrated that TtPaaI can serve as a framework for functional multivalent display of ligands. •We have identified thermostable TtPaaI as potential scaffold for tetramerization of proteins.•We have prepared recombinant fusion between TtPaaI and galectin-7.•We have demonstrated that TtPaaI-galectin-7 is oligomeric and functional.•Our data imply that TtPaaI can be used as scaffold for display of proteins.