scholarly journals Catalytically important damage-free structures of a copper nitrite reductase obtained by femtosecond X-ray laser and room-temperature neutron crystallography

IUCrJ ◽  
2019 ◽  
Vol 6 (4) ◽  
pp. 761-772 ◽  
Author(s):  
Thomas P. Halsted ◽  
Keitaro Yamashita ◽  
Chai C. Gopalasingam ◽  
Rajesh T. Shenoy ◽  
Kunio Hirata ◽  
...  
Keyword(s):  
High Resolution ◽  
Resting State ◽  
Structural Data ◽  
Redox System ◽  
X Rays ◽  
X Ray ◽  
Nitrite Reductases ◽  
First Time ◽  
Better Than ◽  
Neutron Crystallography

Copper-containing nitrite reductases (CuNiRs) that convert NO2−to NO via a CuCAT–His–Cys–CuETproton-coupled redox system are of central importance in nitrogen-based energy metabolism. These metalloenzymes, like all redox enzymes, are very susceptible to radiation damage from the intense synchrotron-radiation X-rays that are used to obtain structures at high resolution. Understanding the chemistry that underpins the enzyme mechanisms in these systems requires resolutions of better than 2 Å. Here, for the first time, the damage-free structure of the resting state of one of the most studied CuNiRs was obtained by combining X-ray free-electron laser (XFEL) and neutron crystallography. This represents the first direct comparison of neutron and XFEL structural data for any protein. In addition, damage-free structures of the reduced and nitrite-bound forms have been obtained to high resolution from cryogenically maintained crystals by XFEL crystallography. It is demonstrated that AspCATand HisCATare deprotonated in the resting state of CuNiRs at pH values close to the optimum for activity. A bridging neutral water (D2O) is positioned with one deuteron directed towards AspCAT Oδ1and one towards HisCAT N∊2. The catalytic T2Cu-ligated water (W1) can clearly be modelled as a neutral D2O molecule as opposed to D3O+or OD−, which have previously been suggested as possible alternatives. The bridging water restricts the movement of the unprotonated AspCATand is too distant to form a hydrogen bond to the O atom of the bound nitrite that interacts with AspCAT. Upon the binding of NO2−a proton is transferred from the bridging water to the Oδ2atom of AspCAT, prompting electron transfer from T1Cu to T2Cu and reducing the catalytic redox centre. This triggers the transfer of a proton from AspCATto the bound nitrite, enabling the reaction to proceed.

scholarly journals IRIXS Spectrograph: an ultra high-resolution spectrometer for tender RIXS

2021 ◽  
Vol 28 (4) ◽  
Author(s):  
Joel Bertinshaw ◽  
Simon Mayer ◽  
Frank-Uwe Dill ◽  
Hakuto Suzuki ◽  
Olaf Leupold ◽  
...  
Keyword(s):  
High Resolution ◽  
Energy Resolution ◽  
X Rays ◽  
Full Width ◽  
X Ray ◽  
X Ray Scattering ◽  
Imaging Approach ◽  
High Resolution Spectrometer ◽  
Resolution Spectrometer ◽  
Better Than

The IRIXS Spectrograph represents a new design of an ultra-high-resolution resonant inelastic X-ray scattering (RIXS) spectrometer that operates at the Ru L 3-edge (2840 eV). First proposed in the field of hard X-rays by Shvyd'ko [(2015), Phys. Rev. A, 91, 053817], the X-ray spectrograph uses a combination of laterally graded multilayer mirrors and collimating/dispersing Ge(111) crystals optics in a novel spectral imaging approach to overcome the energy resolution limitation of a traditional Rowland-type spectrometer [Gretarsson et al. (2020), J. Synchrotron Rad. 27, 538–544]. In combination with a dispersionless nested four-bounce high-resolution monochromator design that utilizes Si(111) and Al2O3(110) crystals, an overall energy resolution better than 35 meV full width at half-maximum has been achieved at the Ru L 3-edge, in excellent agreement with ray-tracing simulations.

Proton Induced X-Ray Fluorescence Analysis Using a Cockcroft-Walton Accelerator

1974 ◽  
Vol 18 ◽  
pp. 299-308
Author(s):  
D. I. Bales ◽  
R. R. Landolt ◽  
W. E. Toll
Keyword(s):  
High Resolution ◽  
Sample Preparation ◽  
Water Sample ◽  
Cross Sections ◽  
Proton Energy ◽  
Fluorescence Analysis ◽  
X Rays ◽  
X Ray ◽  
Beam Currents ◽  
Better Than

AbstractThe production of characteristic x rays by 100-keV protons has been studied to determine the feasibility of using a Cockcroft- Walton accelerator as an excitation source for x ray fluorescence analysis. Because of the low cross sections at this proton energy, it was necessary to concentrate on characteristic x rays below 6 keV and to use beam currents of 10 to 100 μA. The x rays were measured by a high resolution Si(Li) detector.The preparation of various types of samples and the problems that arise when working at low proton energies and high beam currents are discussed.It was found that the technique is useful for low atomic number elements down to less than 1 ppm in a water sample when there are no other significant impurities. When other elements are present, however, proton absorption becomes an important factor and quantitative determinations become very difficult. Proton absorption and its effect upon the usefulness of the technique are discussed.The precision for the technique was determined and found to be better than 20% for most measurements. This includes errors in sample preparation which was considered to be a significant portion of the uncertainty.

Tubulin structure at intermediate resolution

1995 ◽  
Vol 53 ◽  
pp. 852-853
Author(s):  
K. H. Downing ◽  
S. G. Wolf ◽  
E. Nogales
Keyword(s):  
High Resolution ◽  
Structural Data ◽  
Therapeutic Drug ◽  
Zinc Ions ◽  
Two Dimensional ◽  
X Ray Diffraction ◽  
X Ray ◽  
Negative Stain ◽  
Functional Mechanisms ◽  
Tubulin Structure

Microtubules are involved in a host of critical cell activities, many of which involve transport of organelles through the cell. Different sets of microtubules appear to form during the cell cycle for different functions. Knowledge of the structure of tubulin will be necessary in order to understand the various functional mechanisms of microtubule assemble, disassembly, and interaction with other molecules, but tubulin has so far resisted crystallization for x-ray diffraction studies. Fortuitously, in the presence of zinc ions, tubulin also forms two-dimensional, crystalline sheets that are ideally suited for study by electron microscopy. We have refined procedures for forming the sheets and preparing them for EM, and have been able to obtain high-resolution structural data that sheds light on the formation and stabilization of microtubules, and even the interaction with a therapeutic drug.Tubulin sheets had been extensively studied in negative stain, demonstrating that the same protofilament structure was formed in the sheets and microtubules. For high resolution studies, we have found that the sheets embedded in either glucose or tannin diffract to around 3 Å.

scholarly journals Observations of X-rays from Laboratory Sparks in Air at Atmospheric Pressure under Negative Switching Impulse Voltages

Atmosphere ◽  
2019 ◽  
Vol 10 (4) ◽  
pp. 169 ◽  
Author(s):  
Mahbubur Rahman ◽  
Pasan Hettiarachchi ◽  
Vernon Cooray ◽  
Joseph Dwyer ◽  
Vladimir Rakov ◽  
...  
Keyword(s):  
Rise Time ◽  
Atmospheric Pressure ◽  
Opposite Polarity ◽  
X Rays ◽  
X Ray ◽  
Impulse Voltage ◽  
First Time ◽  
Switching Impulses ◽  
Low Rate

We present observations of X-rays from laboratory sparks created in the air at atmospheric pressure by applying an impulse voltage with long (250 µs) rise-time. X-ray production in 35 and 46 cm gaps for three different electrode configurations was studied. The results demonstrate, for the first time, the production of X-rays in gaps subjected to switching impulses. The low rate of rise of the voltage in switching impulses does not significantly reduce the production of X-rays. Additionally, the timing of the X-ray occurrence suggests the possibility that the mechanism of X-ray production by sparks is related to the collision of streamers of opposite polarity.

Mesoscopic structure of SiC fibers by neutron and x-ray scattering

1996 ◽  
Vol 11 (5) ◽  
pp. 1169-1178 ◽  
Author(s):  
Kentaro Suzuya ◽  
Michihiro Furusaka ◽  
Noboru Watanabe ◽  
Makoto Osawa ◽  
Kiyohito Okamura ◽  
...  
Keyword(s):  
Neutron Scattering ◽  
Volume Fraction ◽  
X Rays ◽  
X Ray ◽  
Sic Fibers ◽  
X Ray Scattering ◽  
Mesoscopic Structure ◽  
Angle Scattering ◽  
Momentum Transfer Range ◽  
First Time

Mesoscopic structures of SiC fibers produced from polycarbosilane by different methods were studied by diffraction and small-angle scattering of neutrons and x-rays. Microvoids of a size of 4–10 Å in diameter have been observed for the first time by neutron scattering in a medium momentum transfer range (Q = 0.1–1.0 Å−1). The size and the volume fraction of β–SiC particles were determined for fibers prepared at different heat-treatment temperatures. The results show that wide-angle neutron scattering measurements are especially useful for the study of the mesoscopic structure of multicomponent materials.

scholarly journals Purification, crystallization and preliminary X-ray diffraction analysis of a hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT) fromCoffea canephorainvolved in chlorogenic acid biosynthesis

2012 ◽  
Vol 68 (7) ◽  
pp. 824-828 ◽  
Author(s):  
Laura A. Lallemand ◽  
James G. McCarthy ◽  
Sean McSweeney ◽  
Andrew A. McCarthy
Keyword(s):  
Structural Data ◽  
Coffea Canephora ◽  
Molecular Replacement ◽  
X Ray Diffraction ◽  
X Ray ◽  
Cell Parameters ◽  
Key Enzyme ◽  
Substrate Specifities ◽  
Drop Technique ◽  
Better Than

Chlorogenic acids (CGAs) are a group of soluble phenolic compounds that are produced by a variety of plants, includingCoffea canephora(robusta coffee). The last step in CGA biosynthesis is generally catalysed by a specific hydroxycinnamoyl-CoA quinate hydroxycinnamoyltransferase (HQT), but it can also be catalysed by the more widely distributed hydroxycinnamoyl-CoA shikimate/quinate hydroxycinnamoyltransferase (HCT). Here, the cloning and overexpression of HCT fromC. canephorainEscherichia colias well as its purification and crystallization are presented. Crystals were obtained by the sitting-drop technique at 293 K and X-ray diffraction data were collected on the microfocus beamline ID23-2 at the ESRF. The HCT crystals diffracted to better than 3.0 Å resolution, belonged to space groupP42212 with unit-cell parametersa=b= 116.1,c= 158.9 Å and contained two molecules in the asymmetric unit. The structure was solved by molecular replacement and is currently under refinement. Such structural data are needed to decipher the molecular basis of the substrate specifities of this key enzyme, which belongs to the large plant acyl-CoA-dependent BAHD acyltransferase superfamily.

Cryogenic high-resolution X-ray spectrometers for SR-XRF and microanalysis

1998 ◽  
Vol 5 (3) ◽  
pp. 515-517 ◽  
Author(s):  
M. Frank ◽  
C. A. Mears ◽  
S. E. Labov ◽  
L. J. Hiller ◽  
J. B. le Grand ◽  
...  
Keyword(s):  
High Resolution ◽  
Energy Resolution ◽  
Superconducting Tunnel Junction ◽  
Semiconductor Detectors ◽  
X Rays ◽  
X Ray ◽  
Sr Xrf ◽  
Demonstration Experiment ◽  
Near Future ◽  
Stj Detector

Experimental results are presented obtained with a cryogenically cooled high-resolution X-ray spectrometer based on a 141 × 141 µm Nb-Al-Al2O3-Al-Nb superconducting tunnel junction (STJ) detector in an SR-XRF demonstration experiment. STJ detectors can operate at count rates approaching those of semiconductor detectors while still providing a significantly better energy resolution for soft X-rays. By measuring fluorescence X-rays from samples containing transition metals and low-Z elements, an FWHM energy resolution of 6–15 eV for X-rays in the energy range 180–1100 eV has been obtained. The results show that, in the near future, STJ detectors may prove very useful in XRF and microanalysis applications.

Applications of High-Resolution Powder X-Ray Diffraction

2007 ◽  
Vol 130 ◽  
pp. 7-14 ◽  
Author(s):  
Andrew N. Fitch
Keyword(s):  
High Resolution ◽  
Synchrotron Radiation ◽  
Powder Diffraction ◽  
X Rays ◽  
X Ray Diffraction ◽  
Crystalline Materials ◽  
X Ray ◽  
Structural Characterisation ◽  
Pdf Analysis

The highly-collimated, intense X-rays produced by a synchrotron radiation source can be harnessed to build high-resolution powder diffraction instruments with a wide variety of applications. The general advantages of using synchrotron radiation for powder diffraction are discussed and illustrated with reference to the structural characterisation of crystalline materials, atomic PDF analysis, in-situ and high-throughput studies where the structure is evolving between successive scans, and the measurement of residual strain in engineering components.

HIGH-RESOLUTION PIXE USING BRAGG’S SPECTROMETER

1991 ◽  
Vol 01 (03) ◽  
pp. 251-258 ◽  
Author(s):  
M. TERASAWA
Keyword(s):  
High Resolution ◽  
Heavy Ions ◽  
X Rays ◽  
Light Elements ◽  
Element Analysis ◽  
X Ray ◽  
Peak Intensity ◽  
Moderate Energy ◽  
Practical Analysis ◽  
Wavelength Selectivity

K, L, and M X-rays in the wavelengths between 6Å and 130Å generated by the bombardment of 200 keV protons and other heavy ions were measured by means of a wavelength dispersive Bragg’s spectrometer. The X-ray peak intensity was fairly high in general, while the background was very low. The technique was favorably applied to a practical analysis of several light elements (Be, B, C, N, O, and F). Use of moderate-energy heavy ions considering the wavelength selectivity in X-ray generation was effective for the element analysis. The high-resolution spectrometry in the analytical application of ion-induced X-ray generation was found to be useful for the study of fine electronic structure, e.g. satellite and hypersatellite X-ray study, and of the chemical state of materials.

Understanding the pH-Dependent Reaction Mechanism of a Glycoside Hydrolase Using High-Resolution X-ray and Neutron Crystallography

ACS Catalysis ◽  
2018 ◽  
Vol 8 (9) ◽  
pp. 8058-8069 ◽  
Author(s):  
Zhihong Li ◽  
Xiaoshuai Zhang ◽  
Qingqing Wang ◽  
Chunran Li ◽  
Nianying Zhang ◽  
...  
Keyword(s):  
Reaction Mechanism ◽  
High Resolution ◽  
Glycoside Hydrolase ◽  
X Ray ◽  
Ph Dependent ◽  
Neutron Crystallography
Sign in / Sign up

Export Citation Format

Share Document