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Fluorescence Detection of a Protein‐Bound 2Fe2S Cluster |
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| Authors: | Kevin G Hoff Dr Rochelle Goodlitt Rui Li Christina D Smolke Dr Jonathan J Silberg Dr |
| Affiliation: | 1. Department of Chemical Engineering, California Institute of Technology, 1200 E. California Boulevard, Pasadena, CA 91125 (USA);2. Department of Biochemistry and Cell Biology, Rice University, 6100 Main Street, Houston, TX, 77005 (USA), Fax: (+1)?713‐348‐5154 |
| Abstract: | A fluorescent biosensor is described for 2Fe2S clusters that is composed of green fluorescent protein (GFP) fused to glutaredoxin 2 (Grx2), as illustrated here. 2Fe2S detection is based on the reduction of GFP fluorescence upon the 2Fe2S‐induced dimerization of GFP‐Grx2. This assay is sufficiently sensitive to detect submicromolar changes in 2Fe2S levels, thus making it suitable for high‐throughput measurements of metallocluster degradation and synthesis reactions.  |
| Keywords: | bioinorganic chemistry biosensors biotechnology iron metalloproteins |