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 a number of
proteins, and experimental conditions for structure determination that are incompatible with the
physiological environment. Today, the ultra-short and ultra-bright pulses of
X-ray
free-electron
lasers have made attainable the dream to determine protein
structures
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. With
the perspective to overcome these challenges, we describe here a new methodology that has
the potential to overcome the signal-to-noise-ratio and protein crystallization limits. Using a
multidisciplinary approach, we propose to create ordered, two dimensional protein arrays
with defined orientation attached on a self-assembled-monolayer. We develop a
literature-based flexible toolbox capable of assembling different kinds of proteins on a
functionalized surface and consider using a graphene cover layer that will allow
performing experiments with proteins in physiological conditions