Fluorescence detection of a protein-bound 2Fe2S cluster.
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.
Hoff
Kevin G
KG
Department of Chemical Engineering, California Institute of Technology, 1200 E. California Boulevard, Pasadena, CA 91125, USA. joff@rice.edu
Goodlitt
Rochelle
R
Li
Rui
R
Smolke
Christina D
CD
Silberg
Jonathan J
JJ
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
D054477:Glutaredoxins; D007506:Iron-Sulfur Proteins