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- Title
Monitoring Endocytic Trafficking of Anthrax Lethal Factor by Precise and Quantitative Protein Labeling.
- Authors
Zheng, Siqi; Zhang, Gong; Li, Jie; Chen, Peng R.
- Abstract
Coupling the genetic code expansion technique with bioorthogonal reactions enables precise control over the conjugation site as well as the choice of fluorescent probes during protein labeling. However, the advantages of this strategy over bulky and rigid fluorescent proteins (FPs) remain to be fully explored. Here we applied site-specific bioorthogonal labeling on anthrax lethal factor (LF) to visualize its membrane translocation inside live cells. In contrast to the previously reported FP tags that significantly perturbed LF's membrane trafficking, our precisely and quantitatively labeled LF exhibited an endocytic activity comparable to wild-type LF. This allowed time-lapse imaging of LF's natural translocation process from host cell membrane to cytosol, which revealed molecular details of its virulence mechanism. Our strategy is generally applicable for monitoring intracellular protein membrane translocation that is difficult to access using conventional protein labeling methodologies.
- Subjects
ANTHRAX; BACTERIAL diseases; PROTEINS; BIOMOLECULES; CELL membranes
- Publication
Angewandte Chemie International Edition, 2014, Vol 53, Issue 25, p6449
- ISSN
1433-7851
- Publication type
Article
- DOI
10.1002/anie.201403945