Perspective: A toolbox for protein structure determination in physiological environment through oriented, 2D ordered, site specific immobilization
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...
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doaj-c90ab7148bb84c4b8d6e2ba8d4476f112020-11-24T23:59:52ZengAIP Publishing LLC and ACAStructural Dynamics2329-77782017-07-0144044017044017-910.1063/1.4981224017795SDYPerspective: A toolbox for protein structure determination in physiological environment through oriented, 2D ordered, site specific immobilizationM. Altissimo0M. Kiskinova1R. Mincigrucci2L. Vaccari3C. Guarnaccia4C. Masciovecchio5 Elettra Sincrotrone Trieste, S. S. 14 km 163, 34149 Trieste, Basovizza, Italy Elettra Sincrotrone Trieste, S. S. 14 km 163, 34149 Trieste, Basovizza, Italy Elettra Sincrotrone Trieste, S. S. 14 km 163, 34149 Trieste, Basovizza, Italy Elettra Sincrotrone Trieste, S. S. 14 km 163, 34149 Trieste, Basovizza, Italy International Centre for Genetic Engineering and Biotechnology, Padriciano 99, 34149 Trieste, Italy Elettra Sincrotrone Trieste, S. S. 14 km 163, 34149 Trieste, Basovizza, ItalyRevealing 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.http://dx.doi.org/10.1063/1.4981224 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
M. Altissimo M. Kiskinova R. Mincigrucci L. Vaccari C. Guarnaccia C. Masciovecchio |
spellingShingle |
M. Altissimo M. Kiskinova R. Mincigrucci L. Vaccari C. Guarnaccia C. Masciovecchio Perspective: A toolbox for protein structure determination in physiological environment through oriented, 2D ordered, site specific immobilization Structural Dynamics |
author_facet |
M. Altissimo M. Kiskinova R. Mincigrucci L. Vaccari C. Guarnaccia C. Masciovecchio |
author_sort |
M. Altissimo |
title |
Perspective: A toolbox for protein structure determination in physiological
environment through oriented, 2D ordered, site specific immobilization |
title_short |
Perspective: A toolbox for protein structure determination in physiological
environment through oriented, 2D ordered, site specific immobilization |
title_full |
Perspective: A toolbox for protein structure determination in physiological
environment through oriented, 2D ordered, site specific immobilization |
title_fullStr |
Perspective: A toolbox for protein structure determination in physiological
environment through oriented, 2D ordered, site specific immobilization |
title_full_unstemmed |
Perspective: A toolbox for protein structure determination in physiological
environment through oriented, 2D ordered, site specific immobilization |
title_sort |
perspective: a toolbox for protein structure determination in physiological
environment through oriented, 2d ordered, site specific immobilization |
publisher |
AIP Publishing LLC and ACA |
series |
Structural Dynamics |
issn |
2329-7778 |
publishDate |
2017-07-01 |
description |
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. |
url |
http://dx.doi.org/10.1063/1.4981224 |
work_keys_str_mv |
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