I would export the protein to the oxidative periplasm or alternatively oxidize the cys-cys-bond with Cu.
Best wishes Kornelius On Fri, 8 Feb 2008 10:44:21 -0500 Kendall Nettles <[EMAIL PROTECTED]> wrote: > I'm trying to engineer a disulfide bond into a protein that has several > other cysteines. > > My question is whether there is a crystallization friendly reducing agent > that can be used to prevent oxidation of the free cysteines without breaking > the disulfide? > > Also, can I expect 100% disulfide formation from standard bacterial > expression (assuming good geometry of the cysteines)? > > > Thanks, > > Kendall > -- > Kendall W. Nettles, PhD > Assistant Professor > Department of Cancer Biology > The Scripps Research Institute > 5353 Parkside Dr. > Jupiter Fl 33458 ---------------------------------------------- Kornelius Zeth Max Planck Institute for Developmental Biology Dept. Protein Evolution Spemannstr. 35 72076 Tuebingen, Germany [EMAIL PROTECTED] Tel -49 7071 601 323 Fax -49 7071 601 349
