Solubilizing buried domains of proteins: a self-assembling interface domain from glutathione reductase.

摘要: In the dimeric glutathione reductase (GR) from Escherichia coli, interface domain is largely surrounded by other three domains in each subunit of protein. Subgenes encoding forms have been expressed E. coli and products purified inclusion bodies: INT excised domain, as it found native GR; INTN INTFN are variants carrying exchanges surface residues what would hydrophobic contact regions with neighboring domains. The isolated was to be a soluble folded protein, but mixture dimer at least two species higher molecular weight. latter were believed arise further association via newly exposed unsatisfied regions. variant INTN, normally involved NADPH-binding GR replaced. This partly suppressed aggregation dimers. However, continued high protein concentrations suggested that one site unwanted still present. After four additional amino acid replacements region FAD-binding resulting exhibited no unspecific aggregation, even 3.2 mg/mL.(ABSTRACT TRUNCATED AT 250 WORDS)