Expression Strategies for Recombinant HECT E3 Ligases in Escherichia coli

A comparative analysis of recombinant expression in E. coli of three HECT E3 ligases reveals a consistent preference for lower expression temperatures. Lower temperatures prevent aggregation and misfolding, enhancing the efficiency and solubility of expressed HECT ligases. Isolated HECT domains generally exhibit higher expression success compared to full-length counterparts, offering improved solubility and yields. However, expression levels vary among ligases, necessitating tailored strategies. Future studies may explore full-length HECT-type E3 ligases in covalent complexes with ubiquitin as a potential, generalizable platform for biomedical research.