Expression of N-terminal Cys-protein fragments using an intein refolding strategy

The inclusion body expression and refolding of a pH-sensitive intein fusion protein (Ssp DnaB intein) delivered sufficient quantities of an N-terminal Cys-polypeptide for native chemical ligations. This strategy circumvents premature intein cleavage under expression conditions and allows the expression and purification of proteins with uncertain solubility properties. The expressed protein resembles the C-terminal portion of the amphiphilic immunity protein Im7, which can be ligated to synthetic thioesters to yield synthetic protein analogues for protein folding studies.

An amphiphilic N-terminal Cys-protein fragment for NCL, which undergoes premature cleavage from a pH-sensitive intein fusion under standard conditions, has been isolated by expression and refolding of inclusion bodies.Figure optionsDownload as PowerPoint slide