Synthetic Preparation of Modified Compounds for Fluorescent Click Labeling of Cysteinylated tRNA
Faculty Mentor Information
Dr. Leslie Nickerson (Mentor), Idaho State University; and Dr. Caryn Evilia (Mentor), Idaho State University
Abstract
The enzyme cysteinyl-tRNA synthetase (CysRS) catalyzes the binding of cysteine to tRNA, a crucial step in protein synthesis. Presently, most studies aiming to characterize enzymes like CysRS utilize radiolabel-based assays to quantitate their products. Although alternative assays avoid radiolabeling, they suffer from other drawbacks, namely lengthy and involved procedures. Combining synthetic click chemistry with a fluorophore may provide an alternate route to avoiding radiolabels. To do so, the fluorophore and the cysteinylated tRNA must first be modified to include clickable functional groups. Towards this, an anthracene derivative containing an alkyne functional group attached to a PEG-based linker has been produced. Future work will include utilizing a diazo-transfer reaction to directly incorporate a clickable azide functional group into cysteine as well as performing the labeling click reaction between the modified fluorophore and azidocysteinylated tRNA.
Synthetic Preparation of Modified Compounds for Fluorescent Click Labeling of Cysteinylated tRNA
The enzyme cysteinyl-tRNA synthetase (CysRS) catalyzes the binding of cysteine to tRNA, a crucial step in protein synthesis. Presently, most studies aiming to characterize enzymes like CysRS utilize radiolabel-based assays to quantitate their products. Although alternative assays avoid radiolabeling, they suffer from other drawbacks, namely lengthy and involved procedures. Combining synthetic click chemistry with a fluorophore may provide an alternate route to avoiding radiolabels. To do so, the fluorophore and the cysteinylated tRNA must first be modified to include clickable functional groups. Towards this, an anthracene derivative containing an alkyne functional group attached to a PEG-based linker has been produced. Future work will include utilizing a diazo-transfer reaction to directly incorporate a clickable azide functional group into cysteine as well as performing the labeling click reaction between the modified fluorophore and azidocysteinylated tRNA.