One of the essential characteristics of any tag used in bioscience and medical applications is its size. The larger the label, the more it may affect the studied object, and the more it may distort its behavior. In this paper, using NMR spectroscopy and X-ray crystallography, we have studied the structure of fluorogen-activating protein FAST both in the apo form and in complex with the fluorogen. We showed that significant change in the protein occurs upon interaction with the ligand. While the protein is completely ordered in the complex, its apo form is characterized by higher mobility and disordering of its N-terminus. We used structural information to design the shortened FAST (which we named nanoFAST) by truncating 26 N-terminal residues. Thus, we created the shortest genetically encoded tag among all known fluorescent and fluorogen-activating proteins, which is composed of only 98 amino acids.
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| http://dx.doi.org/10.1039/d1sc01454d | DOI Listing |
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Article Synopsis
- The study explores how fluorescent proteins (FPs) can be used to track proteins in live cells, highlighting limitations when FPs label all protein pools.
- To address this, the research introduces bioconjugate tags known as fluorogen activation proteins (FAPs), which enable selective visualization of specific protein subpopulations.
- The researchers optimized FAP technology for use in yeast, successfully tagging a G-protein coupled receptor to examine its behavior in response to external signals, enhancing the potential applications of FAPs in cellular studies.
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