scholarly journals Crosstalk between Diverse Synthetic Protein Degradation Tags inEscherichia coli

2017 ◽  
Vol 7 (1) ◽  
pp. 54-62 ◽  
Author(s):  
Nicholas C. Butzin ◽  
William H. Mather
Keyword(s):  
Protein Degradation ◽  
Inescherichia Coli ◽  
Synthetic Protein

scholarly journals Crosstalk between diverse synthetic protein degradation tags inEscherichia coli

2017 ◽  
Author(s):  
Nicholas C. Butzin ◽  
William H. Mather
Keyword(s):  
Protein Degradation ◽  
Fluorescent Proteins ◽  
Inescherichia Coli ◽  
E Coli ◽  
Synthetic Protein ◽  
Degradation Rates ◽  
Cell Assays ◽  
Synthetic Circuit ◽  
Microplate Reader ◽  
Single Protease

AbstractRecently, a synthetic circuit inE. colidemonstrated that two proteins engineered with LAA tags targeted to the native protease ClpXP are susceptible to crosstalk due to competition for degradation between proteins. To understand proteolytic crosstalk beyond the single protease regime, we investigated inE. colia set of synthetic circuits designed to probe the dynamics of existing and novel degradation tags fused to fluorescent proteins. These circuits were tested using both microplate reader and single-cell assays. We first quantified the degradation rates of each tag in isolation. We then tested if there was crosstalk between two distinguishable fluorescent proteins engineered with identical or different degradation tags. We demonstrated that proteolytic crosstalk was indeed not limited to the LAA degradation tag, but was also apparent between other diverse tags, supporting the complexity of theE. coliprotein degradation system.

A Caged E3 Ligase Ligand for PROTAC-Mediated Protein Degradation with Light

2019 ◽  
Author(s):  
Cyrille Kounde ◽  
Maria M. Shchepinova ◽  
Edward Tate
Keyword(s):  
Protein Degradation ◽  
Irradiation Time ◽  
E3 Ligase ◽  
Von Hippel Lindau ◽  
Short Irradiation ◽  
Short Irradiation Time ◽  
Spatiotemporal Control

A caging group has been appended to a widely used Von Hippel Lindau (VHL) E3 ligase ligand for targeted protein degradation with PROTACs. Proteolysis is triggered only after a short irradiation time allowing spatiotemporal control of the protein’s fate.

PHOTACs Enable Optical Control of Protein Degradation

2019 ◽  
Author(s):  
Martin Reynders ◽  
Bryan Matsuura ◽  
Marleen Bérouti ◽  
Daniele Simoneschi ◽  
Antonio Marzio ◽  
...  
Keyword(s):  
Protein Degradation ◽  
E3 Ligase ◽  
Optical Control ◽  
Cellular Proteins ◽  
Bifunctional Molecules ◽  
Protein Levels ◽  
Lead Compounds ◽  
Subsequent Degradation ◽  
Temporal And Spatial ◽  
Precision Therapeutics

<p><i>PROTACs (proteolysis targeting chimeras) are bifunctional molecules that tag proteins for ubiquitylation by an E3 ligase complex and subsequent degradation by the proteasome. They have emerged as powerful tools to control the levels of specific cellular proteins and are on the verge of being clinically used. We now introduce photoswitchable PROTACs that can be activated with the temporal and spatial precision that light provides. These trifunctional molecules, which we named PHOTACs, consist of a ligand for an E3 ligase, a photoswitch, and a ligand for a protein of interest. We demonstrate this concept by using PHOTACs that target either BET family proteins (BRD2,3,4) or FKBP12. Our lead compounds display little or no activity in the dark but can be reversibly activated to varying degrees with different wavelengths of light. Our modular and generalizable approach provides a method for the optical control of protein levels with photopharmacology and could lead to new types of precision therapeutics that avoid undesired systemic toxicity.</i><b></b></p>

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