scholarly journals X-ray Collimation for Protein Crystallography

1985 ◽  
Vol 38 (3) ◽  
pp. 353 ◽  
Author(s):  
UW Arndt ◽  
DJ Thomas

Native protein crystals frequently exhibit a very low mosaicity: the mosaic blocks are large and their relative mis-orientation is small. With suitable collimation it is possible to make use of this perfection so as to obtain diffraction profiles with a very narrow width and, accordingly, to improve the spot-to-background ratio and to increase the recorded intensity.

2018 ◽  
Vol 74 (8) ◽  
pp. 792-799 ◽  
Author(s):  
Hanna Kwon ◽  
Patricia S. Langan ◽  
Leighton Coates ◽  
Emma L. Raven ◽  
Peter C. E. Moody

The use of boiled-off liquid nitrogen to maintain protein crystals at 100 K during X-ray data collection has become almost universal. Applying this to neutron protein crystallography offers the opportunity to significantly broaden the scope of biochemical problems that can be addressed, although care must be taken in assuming that direct extrapolation to room temperature is always valid. Here, the history to date of neutron protein cryo-crystallography and the particular problems and solutions associated with the mounting and cryocooling of the larger crystals needed for neutron crystallography are reviewed. Finally, the outlook for further cryogenic neutron studies using existing and future neutron instrumentation is discussed.


Neutrons interact differently with protein crystals than do X-rays. Not only do hydrogen or deuterium atoms diffract neutrons relatively more strongly, but in addition protein crystals suffer no radiation damage in a neutron beam. These and other differences are being exploited for a few selected proteins, at three reactors. At the Institut Laue-Langevin, Grenoble, the crystal structure of the triclinic form of hen egg-white lysozyme is being refined at high resolution ( d -spacings down to 1.4 Å (0.14 nm)), from neutron diffraction measurements on a partly deuterated native crystal of volume 20 mm 3 . Results at the present stage of refinement are discussed. Prospects for neutron protein crystallography are examined in the light of progress in X-ray protein crystallography and neutron detector technology.


Author(s):  
S. M. Zemyan ◽  
D. B. Williams

As has been reported elsewhere, a thin evaporated Cr film can be used to monitor the x-ray peak to background ratio (P/B) in an analytical electron microscope. Presented here are the results of P/B measurements for the Cr Ka line on a Philips EM430 TEM/STEM, with Link Si(Li) and intrinsic Ge (IG) x-ray detectors. The goal of the study was to determine the best conditions for x-ray microanalysis.We used the Fiori P/B definition, in which P/B is the ratio of the total peak integral to the average background in a 10 eV channel beneath the peak. Peak and background integrals were determined by the window method, using a peak window from 5.0 to 5.7 keV about Cr Kα, and background windows from 4.1 to 4.8 keV and 6.3 to 7.0 keV.


Author(s):  
B. B. Chang ◽  
D. F. Parsons

The significance of dynamical scattering effects remains the major question in the structural analysis by electron diffraction of protein crystals preserved in the hydrated state. In the few cases (single layers of purple membrane and 400-600 Å thick catalase crystals examined at 100 kV acceleration voltage) where electron-diffraction patterns were used quantitatively, dynamical scattering effects were considered unimportant on the basis of a comparison with x-ray intensities. The kinematical treatment is usually justified by the thinness of the crystal. A theoretical investigation by Ho et al. using Cowley-Moodie multislice formulation of dynamical scattering theory and cytochrome b5as the test object2 suggests that kinematical analysis of electron diffraction data with 100-keV electrons would not likely be valid for specimen thickness of 300 Å or more. We have chosen to work with electron diffraction patterns obtained from actual wet protein crystals (rat hemoglobin crystals of thickness range 1000 to 2500 Å) at 200 and 1000 kV and to analyze these for dynamical effects.


Author(s):  
Koji INAKA ◽  
Saori ICHIMIZU ◽  
Izumi YOSHIZAKI ◽  
Kiyohito KIHIRA ◽  
Elena G. LAVRENKO ◽  
...  

A series of space experiments aboard the International Space Station (ISS) associated with high-quality Protein Crystal Growth (PCG) in microgravity conditions can be considered as a unique and one of the best examples of fruitful collaboration between Japanese and Russian scientists and engineers in space, which includes also other ISS International Partners. X-ray diffraction is still the most powerful tool to determine the protein three dimensional structure necessary for Structure based drug design (SBDD). The major purpose of the experiment is to grow high quality protein crystals in microgravity for X-ray diffraction on Earth. Within one and a half decade, Japan and Russia have established an efficient process over PCG in space to support latest developments over drug design and structural biology. One of the keys for success of the experiment lies in how precisely pre-launch preparations are made. Japanese party provides flight equipment for crystallization and ensures the required environment to support the experiment aboard of the ISS’s Kibo module, and also mainly takes part of the experiment ground support such as protein sample characterization, purification, crystallization screening, and solution optimization for microgravity experiment. Russian party is responsible for integration of the flight items equipped with proteins and precipitants on board Russian transportation space vehicles (Soyuz or Progress), for delivery them at the ISS, transfer to Kibo module, and returning the experiments’ results back on Earth aboard Soyuz manned capsule. Due to close cooperation of the parties and solid organizational structure, samples can be launched at the ISS every half a year if the ground preparation goes smoothly. The samples are crystallized using counter diffusion method at 20 degree C for 1–2.5 months. After samples return, the crystals are carefully taken out from the capillary, and frozen for X-ray diffraction at SPring8 facility in Japan. Extensive support of researchers from both countries is also a part of this process. The paper analyses details of the PCG experiment scheme, unique and reliable technology of its execution, and contains examples of the application. Key words: International Space Station, Protein crystals, Microgravity, International collaboration.


2018 ◽  
Vol 5 (5) ◽  
pp. 054303 ◽  
Author(s):  
Nadia L. Opara ◽  
Istvan Mohacsi ◽  
Mikako Makita ◽  
Daniel Castano-Diez ◽  
Ana Diaz ◽  
...  
Keyword(s):  

1981 ◽  
Vol 14 (1) ◽  
pp. 33-37 ◽  
Author(s):  
T. D. Mokulskaya ◽  
S. V. Kuzev ◽  
G. E. Myshko ◽  
A. A. Khrenov ◽  
M. A. Mokulskii ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document