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Remdesivir metabolite GS-441524 binds to the SARS-CoV-2 protein nsP3 – potential for drug development to combat numerous other virusesÂ
When a cell is infected, SARS-CoV-2 not only causes
the host cell to produce new virus particles. The virus also suppresses host
cell defence mechanisms. The virus protein nsP3 plays a central role in this.
Using structural analyses, researchers at 51ÁÔÆæ in cooperation with
the Swiss Paul Scherrer Institute have now discovered that a decomposition
product of the virostatic agent remdesivir binds to nsP3. This points to a
further, previously unknown effective mechanism of remdesivir which may be
important for the development of new drugs to combat SARS-CoV-2 and other RNA
viruses.
FRANKFURT. The
virostatic agent remdesivir was developed to disrupt an important step in the
propagation of RNA viruses, to which SARS-CoV-2 also belongs: the reproduction
of the virus's own genetic material. This is present as RNA matrices with which
the host cell directly produces virus proteins. To accelerate the production of
its own proteins, however, RNA viruses cause the RNA matrices to be copied. To
do so, they use a specific protein of their own (an RNA polymerase), which is
blocked by remdesivir. Strictly speaking, remdesivir does not do this itself,
but rather a substance that is synthesized from remdesivir in five steps when
remdesivir penetrates a cell.
In the second of these five steps, an
intermediate is formed from remdesivir, a substance with the somewhat unwieldy
name GS-441524 (in scientific terms: a remdesivir metabolite). GS-441524 is a virostatic
agent as well. As the scientists in the group headed by Professor Stefan Knapp
from the Institute for Pharmaceutical Chemistry at 51ÁÔÆæ Frankfurt
have discovered, GS-441524 targets a SARS-CoV-2 protein called nsP3. nsP3 is a
multifunctional protein, whose tasks include suppressing the host cell's defence
response. The host cell is not helpless in the face of a virus attack, but
activates inflammatory mechanisms, among other things, to summon the aid of the
cell's endogenous immune system. nsP3 helps the viruses suppress the cell's
calls for help.
Professor Stefan Knapp explains: “GS-441525
inhibits the activities of an nsP3 domain which is important for the reproduction
of viruses, and which communicates with human cellular defence systems. Our
structural analysis shows how this inhibition functions, allowing us to lay an
important foundation for the development of new and more potent antiviral drugs
– effective not only against SARS-CoV-2. The target structure of GS-441524 is
very similar in other coronaviruses, for example SARS-CoV and MERS-CoV, as well
in a series of alphaviruses, such as the chikungunya virus. For this reason,
the development of such medicines could also help prepare for future virus
pandemics."
Publication:
Xiaomin Ni, Martin Schröder, Vincent
Olieric, May E. Sharpe, Victor Hernandez-Olmos, Ewgenij Proschak, Daniel Merk,
Stefan Knapp, Apirat Chaikuad: Structural
Insights into Plasticity and Discovery of Remdesivir Metabolite GS-441524
Binding in SARS-CoV‑2 Macrodomain. ACS Med. Chem. Lett. 2021, 12, 603−609
Further
information
Professor Stefan Knapp
Institute for Pharmaceutical Chemistry and
Buchmann Institute for Molecular Life
Sciences
51ÁÔÆæ Frankfurt
Tel. +49 69 798-29871
knapp@pharmchem.uni-frankfurt.de
Editor: Dr. Markus Bernards, Science Editor, PR & Communication Department, Tel: -49 (0) 69 798-12498, Fax: +49 (0) 69 798-763 12531, E-Mail: bernards@em.uni-frankfurt.de
DFG Research Training Group “Configurations of film“ at 51ÁÔÆæ can continue its work
What happens when film
leaves the cinema and becomes available everywhere – out and about on mobile
devices, or in the living room at home? The Graduiertenkolleg (Research
Training Group) “Configurations of Film" at 51ÁÔÆæ has been
researching the current transformation of film and cinema culture since 2017.
The German Research Foundation has now given the project the green light to
continue.
FRANKFURT.
“We are
happy that the German Research Foundation's has kept their trust in us so that
we can continue to our work in the Kolleg," says Vinzenz Hediger, professor for
film studies and speaker of the Kolleg. In individual studies that include the participation
of the disciplines of philosophy, literary studies and theatre studies, the Kolleg
examines a fundamental problem in film studies: the transformation of its
objects through the progressive digitalisation of the production, distribution
and perception of moving images. "The medium of the moving image, which
was standardised for global distribution in an international agreement as early
as 1905, has always been a medium in motion," says Hediger. "With
digitalisation, however, cinema itself as the privileged place of film is now
being called into question, with far-reaching consequences for the aesthetics, as
well as for the social impact and significance of films and other moving image
formats."
The Graduiertenkolleg at
the Institute for Theatre, Film and Media Studies started in 2017 with twelve
doctoral candidates and two post-docs. Currently, the second group with a
another twelve young sceintists from Germany, India and Nigeria is already at
work. In close collaboration with the two postdocs of the Kolleg, they deal
with topics as diverse as the interpenetration of film and video and computer
games, the afterlife of Rainer Werner Fassbinder's work and reputation, the
role of textiles in Nigerian historical films or the digital rediscovery of
popular Bengali cinema of the 1950s and 1960s.
The Graduiertenkolleg is
run in cooperation with the Universities of Mainz and Marburg and the University
for Art and Design in Offenbach. The Kolleg builds on three Master's programmes
at 51ÁÔÆæ as well as collaborations among the applicant researchers.
It utilises the potential of Frankfurt as a location, where the university
library and the German National Library have literature holdings of European
standing and important non-university partners are available in the form of the
German Film Institute, the Murnau Foundation and the Max Planck Institute for
Empirical Aesthetics. The Kolleg is developing an international reputation
through its cooperation with Yale University and Concordia University.
The Kolleg attracted
attention among experts in autumn 2020 with the publication "Pandemic
Media. Preliminary Notes towards an Inventory", in which 37 authors from
the Kolleg and its international network reflect on global media culture under
pandemic conditions. The book is available in open access at the academic
publisher meson press ().
Further
information:
Professor Vinzenz Hediger
Graduiertenkolleg
„Configurations of Film“
Editor: Dr. Anke Sauter, Science and Humanities Editor, International Communication, PR & Communication Department, Phone: +49 69 798-13066, Fax +49(0)69 798-761 12531, sauter@pvw.uni-frankfurt.de.
80 percent of all SARS-CoV-2 proteins produced in the laboratory – protocols available for worldwide research - 51ÁÔÆæ Frankfurt forms the hub of research network from 17 countries
For the development of drugs or vaccines against
COVID-19, research needs virus proteins of high purity. For most of the
SARS-CoV-2 proteins, scientists at 51ÁÔÆæ Frankfurt and a total of
36 partner laboratories have now developed protocols that enable the production
of several milligrams of each of these proteins with high purity, and allow the
determination of the three-dimensional protein structures. The laboratory protocols
and the required genetic tools are freely accessible to researchers all over
the world.
FRANKFURT. When
the SARS-CoV-2 virus mutates, this initially only means that there is a change
in its genetic blueprint. The mutation may lead, for example, to an amino acid
being exchanged at a particular site in a viral protein. In order to quickly
assess the effect of this change, a three-dimensional image of the viral
protein is extremely helpful. This is because it shows whether the switch in amino
acid has consequences for the function of the protein - or for the interaction
with a potential drug or antibody.
Researchers at 51ÁÔÆæ Frankfurt
and TU Darmstadt began networking internationally from the very start of the
pandemic. Their goal: to describe the three-dimensional structures of
SARS-CoV-2 molecules using nuclear magnetic resonance spectroscopy (NMR). In
NMR spectroscopy, molecules are first labelled with special types of atoms
(isotopes) and then exposed to a strong magnetic field. NMR can then be used to
look in detail and with high throughput at how potentially active compounds
bind to viral proteins. This is done at the Centre for Biomolecular Magnetic
Resonance (BMRZ) at 51ÁÔÆæ and other locations. However, the basic
prerequisite is to produce large quantities of the proteins in high purity and
stability, and with their correct folding, for the large amount of tests.
The network, coordinated by Professor
Harald Schwalbe from the Institute of Organic Chemistry and Chemical Biology at
51ÁÔÆæ, spans the globe. The elaboration of laboratory protocols for
the production of proteins is already the second milestone. In addition to
proteins, the virus consists of RNA, and the consortium already made all important RNA fragments of SARS-CoV-2 accessible last year. With the expertise of 129 colleagues,
it has now been possible to produce and purify 23 of the total of almost 30
proteins of SARS-CoV-2 completely or as relevant fragments "in the test
tube", and in large amounts.
For this purpose, the genetic information
for these proteins was incorporated into small, ring-shaped pieces of DNA
(plasmids). These plasmids were then introduced into bacteria for protein
production. Some special proteins were also produced in cell-free systems.
Whether these proteins were still correctly folded after their isolation and
enrichment was confirmed, among other things, by NMR spectroscopy.
Dr Martin Hengesbach from the Institute of
Organic Chemistry and Chemical Biology at 51ÁÔÆæ explains: "We
have isolated functional units of the SARS-CoV-2 proteins in such a way that
their structure, function and interactions can now be characterised by
ourselves and others. In doing so, our large consortium provides working
protocols that will allow laboratories around the world to work quickly and
reproducibly on SARS-CoV-2 proteins and also the mutants to come. Distributing
this work from the beginning was one of our most important priorities. In
addition to the protocols, we are also making the plasmids freely
available."
Dr Andreas Schlundt from the Institute for
Molecular Biosciences at 51ÁÔÆæ says: "With our work, we are
speeding up the global search for active agents: Scientific laboratories equipped
for this work do not have to first spend several months establishing and
optimising systems for the production and investigation of SARS-CoV-2 proteins,
but can now start their research work within two weeks thanks to our elaborated
protocols. Given the numerous mutations of SARS-CoV-2 to come, it is
particularly important to have access to reliable, rapid and well-established
methods for studying the virus in the laboratory. This will, for example, also
facilitate research on the so-called helper proteins of SARS-CoV-2, which have
remained under-investigated, but which also play a role in the occurrence of
mutations."
In the meantime, the work in the NMR
consortium continues: Currently, the researchers are working hard to find out
whether viral proteins can bind to potential drugs.
The research work was funded by the German
Research Foundation and the Goethe Coronavirus Fund. The high logistical effort
and constant communication of research results was supported by Signals, a
spin-off company of 51ÁÔÆæ.
Publication:
Nadide Altincekic, Sophie Marianne Korn,
Nusrat Shahin Qureshi, Marie Dujardin, Martà Ninot-Pedrosa
et. al. Large-scale recombinant
production of the SARS-CoV-2 proteome for high-throughput and structural
biology applications. Frontiers in Molecular Biosciences.
Additional
information: Folding of SARS-CoV2 genome reveals drug
targets – and preparation for “SARS-CoV3"
Images
may be downloaded here: www.uni-frankfurt.de/100668377
Caption:
Scientists Martin
Hengesbach (left) und Andreas Schlundt at the nuclear magnetic resonance (NMR)
spectrometre at Goethe-University Frankfurt, Germany. Photo: Uwe Dettmar for
Goethe-University Frankfurt, Germany
The
COVID-19 NMR Consortium:
Scientific
contacts at 51ÁÔÆæ Frankfurt:
Dr Andreas Schlundt
Emmy Noether Junior Group Leader
Institute for Molecular Biosciences
51ÁÔÆæ Frankfurt
Tel.: +49 69 798-29699
schlundt@bio.uni-frankfurt.de
Dr Martin Hengesbach
Junior Group Leader
51ÁÔÆæ Frankfurt
Institute for Organic Chemistry and Chemical Biology
SFB 902 “Molecular Principles of RNA-based Regulation“
Tel.: +49 69 798-29130
hengesbach@nmr.uni-frankfurt.de
Partners:
Brazil
France
Germany
Greece
Italy
Latvia
Switzerland
Spain
USA
Editor: Dr. Markus Bernards, Science Editor, PR & Communication Department, Tel: -49 (0) 69 798-12498, Fax: +49 (0) 69 798-763 12531, E-Mail: bernards@em.uni-frankfurt.de
51ÁÔÆæ researchers investigate oxidative stress in mice
Oxygen radicals in the body are generally considered
dangerous because they can trigger something called oxidative stress, which is
associated with the development of many chronic diseases such as cancer and
cardiovascular disease. In studies on mice, scientists at 51ÁÔÆæ
Frankfurt have now discovered how oxygen radicals, conversely, can also reduce
the risk of cancer and mitigate damage to the hereditary molecule DNA. (PNAS, DOI 10.1073/pnas.2020152118).
FRANKFURT. Originally,
oxygen radicals - reactive oxygen species, or ROS for short - were considered to
be exclusively harmful in the body. They are produced, for example, by smoking
or UV radiation. Because of their high reactivity, they can damage many
important molecules in cells, including the hereditary molecule DNA. As a
result, there is a risk of inflammatory reactions and the degeneration of
affected cells into cancer cells.
Because of their damaging effect, however,
ROS are also deliberately produced by the body, for example by immune or lung
epithelial cells, which destroy invading bacteria and viruses with ROS. This
requires relatively high ROS concentrations. In low concentrations, on the
other hand, ROS play an important role as signalling molecules. For these
tasks, ROS are specifically produced by a whole group of enzymes. One
representative of this group of enzymes is Nox4, which continuously produces
small amounts of H2O2.
Nox4 is found in almost all body cells, where its product H2O2 maintains a large number of specialised signaling
functions, contributing, for example, to the inhibition of inflammatory
reactions.
Researchers at 51ÁÔÆæ
Frankfurt, led by Professor Katrin Schröder, have now discovered that by
producing H2O2,
Nox4 can even prevent the development of
cancer. They examined mice that were unable to produce Nox4 due to a genetic
modification. When these mice were exposed to a carcinogenic environmental
toxin (cancerogen), the probability that they would develop a tumour doubled.
Since the mice suffered from very different types of tumours such as skin
sarcomas and colon carcinomas, the researchers suspected that Nox4 has a
fundamental influence on cellular health.
Molecular investigations showed that the H2O2
formed by Nox4 keeps a cascade going that prevents certain important signalling
proteins (phosphatases) from entering the cell nucleus. If Nox4 and consequently
H2O2 are absent, those signalling proteins migrate into
the cell nucleus and as a consequence, severe DNA damage is hardly recognised.
Severe DNA damage - e.g. double strand
breaks - occurs somewhere in the body every day. Cells react very sensitively
to such DNA damage, setting a whole repertoire of repair enzymes in motion. If
this does not help, the cell activates its cell death programme - a
precautionary measure of the body against cancer. When such damage goes
unrecognised, as occurs in the absence of Nox4, it spurs cancer formation.
Prof. Katrin Schröder explains the
research results: "If Nox4 is missing and there is therefore no H2O2,
the cells no longer recognise DNA damage. Mutations accumulate and damaged
cells continue to multiply. If an environmental toxin is added that massively
damages the DNA, the damage is no longer recognised and repaired. The affected
cells are not eliminated either, but multiply, sometimes very quickly and
uncontrollably, which eventually leads to the development of tumours. A small
amount of H2O2 thus maintains an internal balance in the
cell that protects the cells from degeneration."
Publication:
Valeska Helfinger, Florian Freiherr von
Gall, Nina Henke, Michael M. Kunze, Tobias Schmid, Flavia Rezende, Juliana
Heidler, Ilka Wittig, Heinfried H. Radeke, Viola Marschall, Karen Anderson,
Ajay M. Shah, Simone Fulda, Bernhard Brüne, Ralf P. Brandes, Katrin Schröder: Genetic deletion of Nox4 enhances
cancerogen-induced formation of solid tumors. PNAS,
Further
information
Professor Katrin Schröder
Institute for Cardiovascular Physiology
Faculty of Medicine
51ÁÔÆæ Frankfurt
Phone +49(0)69-6301-83660
schroeder@vrc.uni-frankfurt.de
Editor: Dr. Markus Bernards, Science Editor, PR & Communication Department, Tel: -49 (0) 69 798-12498, Fax: +49 (0) 69 798-763 12531, E-Mail: bernards@em.uni-frankfurt.de
Scientists at 51ÁÔÆæ and University of Bristol (UK) find traces of beeswax in prehistoric pottery of the West African Nok culture
Before sugar cane and sugar beets conquered the world,
honey was the worldwide most important natural product for sweetening. Archaeologists
at 51ÁÔÆæ in cooperation with chemists at the University of Bristol
have now produced the oldest direct evidence of honey collecting of in Africa.
They used chemical food residues in potsherds found in Nigeria. (Nature
Communications, DOI 10.1038/s41467-021-22425-4)
FRANKFURT. Honey
is humankind's oldest sweetener – and for thousands of years it was also the
only one. Indirect clues about the significance of bees and bee products are provided
by prehistoric petroglyphs on various continents, created between 8,000 and
40,000 years ago. Ancient Egyptian reliefs indicate the practice of beekeeping
as early as 2600 year BCE. But for sub-Saharan Africa, direct archaeological
evidence has been lacking until now. The analysis of the chemical residues of
food in potsherds has fundamentally altered the picture. Archaeologists at Goethe
University in cooperation with chemists at the University of Bristol were able
to identify beeswax residues in 3500 year-old potsherds of the Nok culture.
The Nok culture in central Nigeria dates
between 1500 BCE and the beginning of the Common Era and is known particularly
for its elaborate terracotta sculptures. These sculptures represent the oldest
figurative art in Africa. Until a few years ago, the social context in which
these sculptures had been created was completely unknown. In a project funded
by the German Research Foundation, 51ÁÔÆæ scientists have been
studying the Nok culture in all its archaeological facets for over twelve
years. In addition to settlement pattern, chronology and meaning of the
terracotta sculptures, the research also focussed on environment, subsistence and
diet.
Did the people of the Nok Culture have domesticated
animals or were they hunters? Archaeologists typically use animal bones from
excavations to answer these questions. But what to do if the soil is so acidic
that bones are not preserved, as is the case in the Nok region?
The analysis of molecular food residues in
pottery opens up new possibilities. This is because the processing of plant and
animal products in clay pots releases stable chemical compounds, especially
fatty acids (lipids). These can be preserved in the pores of the vessel walls
for thousands of years, and can be detected with the assistance of gas
chromatography.
To the researchers' great surprise, they
found numerous other components besides the remains of wild animals, significantly
expanding the previously known spectrum of animals and plants used. There is
one creature in particular that they had not expected: the honeybee. A third of
the examined shards contained high-molecular lipids, typical for beeswax.
It is not possible to reconstruct from the
lipids which bee products were used by the people of the Nok culture. Most probably
they separated the honey from the waxy combs by heating them in the pots. But
it is also conceivable that honey was processed together with other raw materials
from animals or plants, or that they made mead. The wax itself could have
served technical or medical purposes. Another possibility is the use of clay
pots as beehives, as is practised to this day in traditional African societies.
“We began this study with our colleagues in
Bristol because we wanted to know if the Nok people had domesticated animals,"
explains Professor Peter Breunig from 51ÁÔÆæ, who is the director of
the archaeological Nok project. “That honey was part of their daily menu was
completely unexpected, and unique in the early history of Africa until now."
Dr Julie Dunne from the University of
Bristol, first author of the study says: “This is a remarkable example for how
biomolecular information from prehistoric pottery in combination with
ethnographic data provides insight into the use of honey 3500 years ago."
Professor Richard Evershed, Head of the
Institute for Organic Chemistry at the University of Bristol and co-author of
the study points out that the special relationship between humans and honeybees
was already known in antiquity. “But the discovery of beeswax residues in Nok
pottery allows a very unique insight into this relationship, when all other
sources of evidence are lacking."
Professor Katharina Neumann, who is in
charge of archaeobotany in the Nok project at 51ÁÔÆæ says: “Plant
and animal residues from archaeological excavations reflect only a small
section of what prehistoric people ate. The chemical residues make previously
invisible components of the prehistoric diet visible." The first direct
evidence of beeswax opens up fascinating perspectives for the archaeology of
Africa. Neumann: “We assume that the use of honey in Africa has a very long
tradition. The oldest pottery on the continent is about 11,000 years old. Does
it perhaps also contain beeswax residues? Archives around the world store thousands
of ceramic shards from archaeological excavations that are just waiting to
reveal their secrets through gas chromatography and paint a picture of the
daily life and diet of prehistoric people."
Publication:
Julie Dunne, Alexa Höhn, Gabriele Franke,
Katharina Neumann, Peter Breunig, Toby Gillard, Caitlin Walton-Doyle, Richard
P. Evershed. Honey-collecting in
prehistoric West Africa from 3500 years ago. Nature Communications
Images
for download:
Traces of beeswax were detected in 3500
year-old clay pots like this (photo: Peter Breunig, 51ÁÔÆæ
Frankfurt)
Dr Gabriele Franke, 51ÁÔÆæ
archaeologist during the documentation of excavated clay pots at the Nok
research station in Janjala, Nigeria in August 2016. Traces of beeswax were
detected in clay pots like these (photo: Peter Breunig, 51ÁÔÆæ
Frankfurt)
Still popular today: excavation workers
enjoy freshly collected wild honey (photo: Peter Breunig, 51ÁÔÆæ
Frankfurt)
The Nok culture is known in Nigeria today for its terracotta figurines (photo: Peter Breunig, 51ÁÔÆæ Frankfurt)
Further
information:
Professor Katharina Neumann
Institute for Archaeological Sciences
51ÁÔÆæ Frankfurt
Phone: 069 798-32292
k.neumann@em.uni-frankfurt.de