Optimization of Copper-Catalyzed Protein Labeling in Biological Systems
Activity-Based Protein Profiling (ABPP) is a chemical biology method to measure activity levels of specific enzymes, or to assist in assigning function to unannotated proteins. The novelty of ABPP is its use of small-molecule probes that can be applied to living cells or cell lysates to selectively label distinct protein families. The probe-labeled proteins can then be isolated from the complex proteome by way of gel electrophoresis or mass spectrometry, and subsequently characterized and quantified. For this project, the Copper-Catalyzed Azide-Alkyne Cycloaddition (CuAAC) that facilitates identification of probe-labeled proteins was optimized. Cyanobacterium Synechococcus sp. PCC 7002 lysates and mouse microsomal fractions served as biological systems for CuAAC testing. CuAAC parameters tested include labeling temperatures, chemical reagents, and reagent ratios. Optimization of the CuAAC reaction within the confines of a biological system increases the efficiency of ABPP by providing improved readouts.
Guerrero, Christian, "Optimization of Copper-Catalyzed Protein Labeling in Biological Systems" (2014). College of Arts and Sciences Presentations. 6.