AI Article Synopsis
- Mapping protein interactions in cells is crucial for understanding their complexities over relevant time periods.
- Researchers developed multicolored luciferase-based systems that allow simultaneous observation of two interacting proteins in live cells using a shared substrate.
- This method was applied to study β-TrCP and its interactions, leading to the identification of GSK3β as a regulating kinase for IκBα processing.
Article Abstract
Understanding the functional complexity of protein interactions requires mapping biomolecular complexes within the cellular environment over biologically relevant time scales. Herein, we describe a set of reversible multicolored heteroprotein complementation fragments based on various firefly and click beetle luciferases that utilize the same substrate, D-luciferin. Luciferase heteroprotein fragment complementation systems enabled dual-color quantification of two discrete pairs of interacting proteins simultaneously or two distinct proteins interacting with a third shared protein in live cells. Using real-time analysis of click beetle green and click beetle red luciferase heteroprotein fragment complementation applied to β-TrCP, an E3-ligase common to the regulation of both β-catenin and IκBα, GSK3β was identified as a candidate kinase regulating IκBα processing. These dual-color protein interaction switches may enable directed dynamic analysis of a variety of protein interactions in living cells.
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| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2943495 | PMC |
| http://dx.doi.org/10.1016/j.chembiol.2010.06.018 | DOI Listing |
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