Work

Light-Activated Proteomic Labeling via Photocaged Bioorthogonal Non-Canonical Amino Acids

Year: 2018

Type: article

Abstract: This work introduces light-activated bioorthogonal noncanonical amino acid tagging (laBONCAT) as a method to selectively label, isolate, and identify proteins newly synthesized at user-defined regions in tissue culture. By photocaging l-azidohomoalanine (Aha), metabolic incorporation into proteins is prevented. The caged compound remains stable for many hours in culture, but can be photochemically liberated rapidly and on demand with spatial control. Upon directed light exposure, the uncaged amino acid is available for local translation, enabling downstream proteomic interrogation via bioorthogonal conjugation. Exploiting the reactive azide moiety present on Aha's amino acid side chain, we demonstrate that newly synthesized proteins can be purified for quantitative proteomics or visualized in synthetic tissues with a new level of spatiotemporal control. Shedding light on when and where proteins are translated within living samples, we anticipate that laBONCAT will aid in understanding the progression of complex protein-related disorders.

Source: ACS Chemical Biology

DOI: 10.1021/acschembio.7b01023

Authors: Steven M. Adelmund, Emily R. Ruskowitz, Payam E. Farahani, Julie V. Wolfe, Cole A. DeForest

Institution: University of Washington

Language: English

Topic: Click Chemistry and Applications

Subfield: Organic Chemistry

Field: Chemistry

Domain: Physical Sciences

Open Access status: green

Funders: National Aeronautics and Space Administration, Washington Research Foundation, University of Washington, National Institutes of Health

Awards: G238994939, ESTABLISHED IN 1989, WASHINGTON NASA SPACE GRANT CONSORTIUM WAS ONE OF THE FIRST SPACE GRANT CONSORTIUMS IN THE COUNTRY AND IS FUNDED BY NASA AS A DE, Leica SP8 WWL Confocal Microscope