Accedi

GenNext Technologies Inc.

3 ARTICLES PUBLISHED IN JoVE

Biochemistry

Characterizing Cellular Proteins with In-cell Fast Photochemical Oxidation of Proteins

Emily E. Chea¹, Aimee Rinas², Jessica A. Espino¹, Lisa M. Jones¹

¹Department of Pharmaceutical Sciences, University of Maryland Baltimore, ²AIT Bioscience

Here, we characterize protein structure and interaction sites in living cells using a protein footprinting technique termed in-cell fast photochemical oxidation of proteins (IC-FPOP).

Biochemistry

Enabling Real-Time Compensation in Fast Photochemical Oxidations of Proteins for the Determination of Protein Topography Changes

Sandeep K. Misra¹, Joshua S. Sharp^1,2,3

¹Department of Biomolecular Sciences, University of Mississippi, ²Department of Chemistry and Biochemistry, University of Mississippi, ³GenNext Technologies, Inc.

Fast photochemical oxidation of proteins is an emerging technique for the structural characterization of proteins. Different solvent additives and ligands have varied hydroxyl radical scavenging properties. To compare the protein structure in different conditions, real-time compensation of hydroxyl radicals generated in the reaction is required to normalize reaction conditions.

JoVE Journal

Laser-free Hydroxyl Radical Protein Footprinting to Perform Higher Order Structural Analysis of Proteins

Scot R. Weinberger¹, Emily E. Chea¹, Joshua S. Sharp^1,2,3, Sandeep K. Misra²

¹GenNext Technologies Inc., ²Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, ³Department of Chemistry and Biochemistry, University of Mississippi

This protocol presents a method to use inline radical dosimetry and a plasma light source to perform flash oxidation protein footprinting. This method replaces the hazardous UV laser to simplify and improve the reproducibility of fast photochemical oxidation of protein studies.