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@INPROCEEDINGS{Schrader:972033,
author = {Schrader, Tobias Erich},
title = {{P}rotein structures and equilibrium dynamics as seen by
neutrons},
reportid = {FZJ-2023-01042},
year = {2022},
abstract = {In this lecture, an introduction into the method of neutron
protein crystallography will be given and the differences to
x-ray crystallography will be highlighted: As opposed to
x-rays, neutrons are scattered from the nuclei and can
therefore locate hydrogen atoms (see Figure 1). Therefore,
typical scientific questions addressed are the determination
of protonation states of amino acid side chains in proteins
and the characterization of the hydrogen bonding networks
between the protein active centre and an inhibitor or
substrate. The neutron single crystal diffractometer BIODIFF
will serve as an example of a neutron protein
crystallography beam line. It is located at the Heinz
Maier-Leibnitz Zentrum, MLZ, at the research reactor (FRM
II) in Garching, Germany. BIODIFF is a joint project of the
Jülich Centre for Neutron Science (JCNS) and the Technical
University of Munich (TUM). BIODFF is equipped with a
standard Oxford Cryosystem “Cryostream 700+” which
allows measurements in the temperature range from 90 K up to
500 K. A new kappa goniometer head was added recently. This
allows an automated tilting of the crystal in order to
increase the completeness of the data set when recording
another set of frames in the tilted geometry. Recently, a
new collimation device was added in front of the detector.
This allows to align the apertures included in the
collimation with a hexapod in all necessary degrees of
freedom. Efforts to increase the flux at the sample position
and to reduce the background at the detector have led to the
ability to measure smaller and smaller protein crystals down
to 0.1 mm3 in volume. One application example is the
improvement of antibiotic drugs. Many bacteria secret a
protein called -lactamase into their environment. This
protein is able to hydrolyse the four membered carbon atom
ring in -lactam antibiotics. These antibiotics are
thereby destroyed and are not harmful to the bacteria any
more. This mechanism causes great problems in hospitals.
With neutron protein crystallography we were able to find a
deuterium atom at the amino acid side chain glutamate 166 in
the -lactamase protein carrying a transition state
analogue. This transition state analogue stops the enzymatic
reaction in its first acylation step. Thereby one could
identify glutamate 166 as the important base taking over the
hydrogen atom in the acylation step. Improved antibiotics
should find ways to bind to this side chain in order to
prevent its action as a base. Or, an additional drug has to
be given to the patients which blocks the -lactamase
protein efficiently such that the antibiotics can work
effectively again. The technique of neutron protein
crystallography uses elastic neutron scattering and gives
information on the structure of the protein. Inelastic
neutron scattering reports on the equilibrium dynamics of
proteins in solution. In a short excursion, neutron spin
echo spectroscopy, an example of an inelastic, i. e.
spectroscopic neutron scattering technique will be
introduced which allows to monitor large scale protein
motions on a nanosecond timescale. In case of the protein
Phosphoglyceratkinase, it will be shown that those motions
are necessary for the protein to fulfill its enzymatic
function. Time permitting, the concept of contrast matching
between solvent and some part of the solute will be
explained and its use in the techniques of small angle
neutron scattering (SANS) and neutron reflectometry will be
discussed briefly.},
month = {Jun},
date = {2022-06-19},
organization = {(FEBS course: Biomolecules in Action
III), 19 Jun 2022 - 24 Jun 2022},
subtyp = {Invited},
cin = {JCNS-FRM-II / MLZ / JCNS-1 / JCNS-4},
cid = {I:(DE-Juel1)JCNS-FRM-II-20110218 / I:(DE-588b)4597118-3 /
I:(DE-Juel1)JCNS-1-20110106 / I:(DE-Juel1)JCNS-4-20201012},
pnm = {6G4 - Jülich Centre for Neutron Research (JCNS) (FZJ)
(POF4-6G4) / 632 - Materials – Quantum, Complex and
Functional Materials (POF4-632)},
pid = {G:(DE-HGF)POF4-6G4 / G:(DE-HGF)POF4-632},
experiment = {EXP:(DE-MLZ)BIODIFF-20140101 / EXP:(DE-MLZ)J-NSE-20140101},
typ = {PUB:(DE-HGF)31},
url = {https://juser.fz-juelich.de/record/972033},
}