A New Route for the Determination of Protein Structure in Physiological Environment

TL;DR

This paper introduces a novel methodology utilizing a 2D protein array on a functionalized surface with a graphene cover, aiming to determine protein structures in physiological environments without crystallization or radiation damage.

Contribution

It proposes a new multidisciplinary approach to overcome limitations of existing techniques by creating oriented protein arrays under physiological conditions.

Findings

Potential to determine protein structures without crystallization

Method preserves proteins in physiological conditions

Addresses radiation damage issues in structure determination

Abstract

Revealing the structure of complex biological macromolecules, such as proteins, is an essential step for understanding the chemical mechanisms that determine the diversity of their functions. Synchrotron based x-ray crystallography and cryo-electron microscopy have made major contributions in determining thousands of protein structures even from micro-sized crystals. They suffer from some limitations that have not been overcome, such as radiation damage, the natural inability to crystallize of a number of proteins and experimental conditions for structure determination that are incompatible with the physiological environment. Today the ultrashort and ultra-bright pulses of X-ray free-electron lasers (XFELs) have made attainable the dream to determine protein structure before radiation damage starts to destroy the samples. However, the signal-to-noise ratio remains a great challenge to obtain usable diffraction patterns from a single protein molecule. We describe here a new methodology that should overcome the signal and protein crystallization limits. Using a multidisciplinary approach, we propose to create a two dimensional protein array with defined orientation attached on a self-assembled-monolayer. We develop a literature-based, flexible toolbox capable of assembling different proteins on a functionalized surface while keeping them under physiological conditions during the experiment, using a water-confining graphene cover.