title: Fluorescence detection of a protein-bound 2Fe2S cluster.
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.
authors.affiliation: Department of Chemical Engineering, California Institute of Technology, 1200 E. California Boulevard, Pasadena, CA 91125, USA. joff@rice.edu
authors.forename: Kevin G
authors.initials: KG
authors.lastname: Hoff
authors.forename: Rochelle
authors.initials: R
authors.lastname: Goodlitt
authors.forename: Rui
authors.initials: R
authors.lastname: Li
authors.forename: Christina D
authors.initials: CD
authors.lastname: Smolke
authors.forename: Jonathan J
authors.initials: JJ
authors.lastname: Silberg
mesh_terms: D015374:Biosensing Techniques / Q000379:methods*; D005453:Fluorescence; D054477:Glutaredoxins / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D007506:Iron-Sulfur Proteins / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D008958:Models, Molecular; D055503:Protein Multimerization; D020836:Protein Structure, Quaternary
chemical_list: D054477:Glutaredoxins; D007506:Iron-Sulfur Proteins