Issue 7, 2015

Protein footprinting by pyrite shrink-wrap laminate

Abstract

The structure of macromolecules and their complexes dictate their biological function. In “footprinting”, the solvent accessibility of the residues that constitute proteins, DNA and RNA can be determined from their reactivity to an exogenous reagent such as the hydroxyl radical (·OH). While ·OH generation for protein footprinting is achieved by radiolysis, photolysis and electrochemistry, we present a simpler solution. A thin film of pyrite (cubic FeS2) nanocrystals deposited onto a shape memory polymer (commodity shrink-wrap film) generates sufficient ·OH via Fenton chemistry for oxidative footprinting analysis of proteins. We demonstrate that varying either time or H2O2 concentration yields the required ·OH dose–oxidation response relationship. A simple and scalable sample handling protocol is enabled by thermoforming the “pyrite shrink-wrap laminate” into a standard microtiter plate format. The low cost and malleability of the laminate facilitates its integration into high throughput screening and microfluidic devices.

Graphical abstract: Protein footprinting by pyrite shrink-wrap laminate

Supplementary files

Article information

Article type
Technical Innovation
Submitted
30 Oct 2014
Accepted
26 Jan 2015
First published
26 Jan 2015

Lab Chip, 2015,15, 1646-1650

Author version available

Protein footprinting by pyrite shrink-wrap laminate

M. Leser, J. Pegan, M. El Makkaoui, J. C. Schlatterer, M. Khine, M. Law and M. Brenowitz, Lab Chip, 2015, 15, 1646 DOI: 10.1039/C4LC01288G

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