Light-Activated Caged Amino Acids for Proteomics Analysis

Principal Investigator: Cole DeForest

Gene expression and protein translation are biological functions that govern a wide range of physiological processes including development, immune behavior, and chemical signaling phenomena. Current methods for analyzing protein production, activity, and identity have mostly centered around mass spectrometry and genomic sequencing, which are limited to endpoint analyses and do not reflect the rapid dynamics associated with protein function. Alternative techniques, such as biorthogonal noncanonical amino acid tagging (BONCAT), enable temporal exploration of protein function through labeling of proteins at specific time points and subsequent monitoring. However, BONCAT does not enable spatial control of protein labeling and analysis.

Researchers in the DeForest Lab have developed laBONCAT (light-activated BONCAT), in which light is used to specifically initiate amino acid uptake for protein fabrication at user-defined times and locations. To enable this precise control in amino acid uptake, they utilize photocages bound to amino acid terminals that block uptake into newly generated proteins. Upon addition of light, the photocage is released and the specific amino acid can be incorporated. Further labeling of these newly translated proteins allows downstream proteomic analysis.

Advantages 

• Temporal and spatial control over availability of amino acids for incorporation by proteins 

• Addition of biorthogonal handles for purification and visualization 

• Theoretically applicable to any amino acid 

• Wide range of available photocages/light exposures can be used


For more info, contact: Ryan Buckmaster

  • swap_vertical_circlelibrary_booksReferences (1)
    1. Steven M. Adelmund, Emily R. Ruskowitz, Payam E. Farahani, Julie V. Wolfe, Cole A. DeForest (2018), Light-Activated Proteomic Labeling via Photocaged Bioorthogonal Non-Canonical Amino Acids, ACS Chem. Biol., 573 - 577
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