I *believe* that the ISC operon is utilized in E. coli for all FeS clusters, regardless if Rieske or not. I know that when deleted, activity of FeS containing proteins (succinate dehydrogenase and Glutamate synthase) greatly decreases. See: http://www.ncbi.nlm.nih.gov/pubmed/11432781
Thus it *should* work with other FeS containing enzymes. Kelly ******************************************************* 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 9:43 AM, Edward A. Berry <ber...@upstate.edu> wrote: > 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<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 >> >> >> >
