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@MISC{Schrader:873038,
author = {Schrader, Tobias Erich},
title = {{N}eutron {P}rotein {C}rystallography and equilibrium
dynamics},
reportid = {FZJ-2020-00489},
year = {2019},
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. 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. 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
agian. 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.},
month = {Jun},
date = {2019-06-25},
organization = {FEBS Practical Course, Biomolecules in
Action II ,DESY, Hamburg (Germany), 25
Jun 2019 - 25 Jun 2019},
subtyp = {Invited},
cin = {JCNS-FRM-II / JCNS-1 / MLZ},
cid = {I:(DE-Juel1)JCNS-FRM-II-20110218 /
I:(DE-Juel1)JCNS-1-20110106 / I:(DE-588b)4597118-3},
pnm = {6G4 - Jülich Centre for Neutron Research (JCNS) (POF3-623)
/ 6G15 - FRM II / MLZ (POF3-6G15) / 6215 - Soft Matter,
Health and Life Sciences (POF3-621)},
pid = {G:(DE-HGF)POF3-6G4 / G:(DE-HGF)POF3-6G15 /
G:(DE-HGF)POF3-6215},
experiment = {EXP:(DE-MLZ)BIODIFF-20140101 / EXP:(DE-MLZ)J-NSE-20140101},
typ = {PUB:(DE-HGF)17},
url = {https://juser.fz-juelich.de/record/873038},
}