Good to know about this ISC operon! Do you know if it is specific for Rieske-type His2/Cys2 Fe2S2 clusters, or for FeS clusters in general? Thus wild-type E. coli is already making three types of ISC clusters for succinate dehydrogenase or fumarate reductase (although some help might be needed for overexpression) but I don't know if it has any Rieske-type clusters.
Kelly Daughtry wrote:
Jan, Do you express the protein with the /E. coli isc/ iron-sulfur cluster synthetic operon? I found great success, see: Daughtry, KD et. al. JACS 2012 http://pubs.acs.org/doi/full/10.1021/ja2111898 - Kelly Daughtry ******************************************************* Kelly Daughtry, Ph.D. Post-Doctoral Fellow, Raetz Lab Biochemistry Department Duke University Alex H. Sands, Jr. Building 303 Research Drive RM 250 Durham, NC 27710 P: 919-684-5178 ******************************************************* On Wed, Jul 11, 2012 at 6:22 AM, Jan Rashid Umar <jan...@googlemail.com <mailto:jan...@googlemail.com>> wrote: Dear All, I will be grateful to your suggestions about Iron-Sulfur cluster protein purification. My colleague has some problems with the purification and it seems that the iron-sulfur cluster might be deformed, and protein is aggregated. The purification is done under aerobic (normal condition), and is there some method to reintegrate iron sulfur cluster back into the protein molecule under these conditions. Can anybody suggest some literature, protocol or something that can improve the protein aggregation? I look forward to hearing from you. Best wishes, Jan
