Monitoring protein stability and aggregation in vivo by real-time fluorescent labeling.

@article{Ignatova2004MonitoringPS,
  title={Monitoring protein stability and aggregation in vivo by real-time fluorescent labeling.},
  author={Zoya Ignatova and Lila M. Gierasch},
  journal={Proceedings of the National Academy of Sciences of the United States of America},
  year={2004},
  volume={101 2},
  pages={523-8}
}
In vivo fluorescent labeling of an expressed protein has enabled the observation of its stability and aggregation directly in bacterial cells. Mammalian cellular retinoic acid-binding protein I (CRABP I) was mutated to incorporate in a surface-exposed omega loop the sequence Cys-Cys-Gly-Pro-Cys-Cys, which binds specifically to a biarsenical fluorescein dye (FlAsH). Unfolding of labeled tetra-Cys CRABP I is accompanied by enhancement of FlAsH fluorescence, which made it possible to determine the… CONTINUE READING
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