The laue method and its use in time-resolved crystallography

1989 ◽  
pp. 61-74 ◽  
Author(s):  
Keith Moffat ◽  
John R. Helliwell
Keyword(s):  
Time Resolved ◽  
Laue Method

A flow cell suitable for time-resolved X-ray crystallography by the Laue method

1997 ◽  
Vol 30 (5) ◽  
pp. 555-556 ◽  
Author(s):  
G. Kurisu ◽  
A. Sugimoto ◽  
Y. Kai ◽  
S. Harada
Keyword(s):  
Flow Cell ◽  
Time Resolved ◽  
X Ray ◽  
X Ray Crystallography ◽  
Laue Method

Time-resolved crystallography on H-ras p21

1992 ◽  
Vol 340 (1657) ◽  
pp. 263-272 ◽  
Keyword(s):  
Active Site ◽  
Protein Product ◽  
X Ray Diffraction ◽  
Time Resolved ◽  
Synchrotron Source ◽  
Current State ◽  
Single Diastereomer ◽  
Ras P21 ◽  
Hydrolysis Of ◽  
Laue Method

We describe here the results obtained to date on a project aimed at characterizing the changes occurring in the protein product (p21) of the H-ras proto-oncogene during and as a result of hydrolysis of GTP at its active site. The approach used involves crystallization of p21 with a photosensitive precursor of GTP (caged GTP) at the active site followed by generation of GTP by photolysis and collection of X-ray diffraction data using the Laue method at a synchrotron source. The structure of p21 complexed with a single diastereomer of caged GTP is presented here. In contrast to crystals obtained with mixed diastereomers, the nucleotide appears to bind in a manner which is very similar to that of other guanine nucleotides (GDP, GTP, GppNHp). The current state of time resolved structural experiments using these crystals is presented.

Time-resolved macromolecular crystallography: introductory remarks and a little history

1992 ◽  
Vol 340 (1657) ◽  
pp. 169-173 ◽  
Keyword(s):  
Crystallographic Data ◽  
Macromolecular Crystallography ◽  
Time Resolved ◽  
X Ray ◽  
Early Use ◽  
Laue Method

X -ray crystallographic data can be collected rapidly at synchrotrons by the Laue method. The early use of the Laue method by Ewald, Nishikawa, Wyckoff and Pauling is reviewed.

The emergence of the synchrotron Laue method for rapid data collection from protein crystals

1993 ◽  
Vol 442 (1914) ◽  
pp. 177-192 ◽  
Keyword(s):  
Multiplicity Distribution ◽  
Spectral Curve ◽  
Three Dimensional ◽  
Protein Crystal ◽  
Photographic Film ◽  
Macromolecular Crystallography ◽  
Time Resolved ◽  
X Ray ◽  
Laue Geometry ◽  
Laue Method

Synchrotron X-radiation (SR) is intense, polychromatic and collimated. It is widely exploited, in macromolecular crystallography, particularly using a monochromatized short wavelength beam. The spectral curve of SR, however, ideally lends itself to use of Laue geometry, i. e. the original diffraction experimental arrangement based on a stationary crystal and a polychromatic X-ray beam. Rapid exposure times and time-resolved crystallography studies, e. g. of enzymes, are now possible. Historical objections to the use of Laue diffraction data, particularly the multiplicity distribution, have been found not to be as limiting as once thought. The credentials of the Laue method have been established through a variety of Laue crystal structure analyses, involving photographic film as detector. Recently a three-dimensional arrangement of films, known as a toast-rack, has been used to alleviate problems with spatially overlapping spots. This paper provides a review of these results and then reports several developments. In particular, one of the first Laue analyses using an image plate as detector, namely of a cobalt substituted concanavalin A crystal, is discussed. Recent experimental developments, also at the Daresbury synchrotron, are then described. First, a large toast-rack has been used to record Laue data from a protein crystal. Secondly, a transmission X-ray mirror has been constructed from thin mylar (1.5 μm) and used to provide a λ max filter instead of using aluminium foils. Thirdly, since the Laue method suffers from poor sampling of the low resolution data, a new method (known as LOT) has been introduced.

Microtubule nucleation sequence studied by time-resolved x-ray scattering and electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 766-767
Author(s):  
Eva-Maria Mandelkow ◽  
Eckhard Mandelkow ◽  
Joan Bordas
Keyword(s):  
Electron Microscopy ◽  
Dynamic Equilibrium ◽  
Time Resolved ◽  
X Ray ◽  
Additional Information ◽  
X Ray Scattering ◽  
Low Concentrations ◽  
Microtubule Growth ◽  
Ray Patterns ◽  
Ray Scattering

When a solution of microtubule protein is changed from non-polymerising to polymerising conditions (e.g. by temperature jump or mixing with GTP) there is a series of structural transitions preceding microtubule growth. These have been detected by time-resolved X-ray scattering using synchrotron radiation, and they may be classified into pre-nucleation and nucleation events. X-ray patterns are good indicators for the average behavior of the particles in solution, but they are difficult to interpret unless additional information on their structure is available. We therefore studied the assembly process by electron microscopy under conditions approaching those of the X-ray experiment. There are two difficulties in the EM approach: One is that the particles important for assembly are usually small and not very regular and therefore tend to be overlooked. Secondly EM specimens require low concentrations which favor disassembly of the particles one wants to observe since there is a dynamic equilibrium between polymers and subunits.

Electron Channeling Patterns

1977 ◽  
Vol 35 ◽  
pp. 12-13
Author(s):  
David C. Joy
Keyword(s):  
Electron Diffraction ◽  
Incidence Angle ◽  
Photographic Plate ◽  
Diffraction Effect ◽  
Angle Of Incidence ◽  
Bulk Single Crystal ◽  
Time Resolved ◽  
Electron Channeling ◽  
Spatially Resolved ◽  
Large Bulk

Electron channeling patterns (ECP) were first found by Coates (1967) while observing a large bulk, single crystal of silicon in a scanning electron microscope. The geometric pattern visible was shown to be produced as a result of the changes in the angle of incidence, between the beam and the specimen surface normal, which occur when the sample is examined at low magnification (Booker, Shaw, Whelan and Hirsch 1967).A conventional electron diffraction pattern consists of an angularly resolved intensity distribution in space which may be directly viewed on a fluorescent screen or recorded on a photographic plate. An ECP, on the other hand, is produced as the result of changes in the signal collected by a suitable electron detector as the incidence angle is varied. If an integrating detector is used, or if the beam traverses the surface at a fixed angle, then no channeling contrast will be observed. The ECP is thus a time resolved electron diffraction effect. It can therefore be related to spatially resolved diffraction phenomena by an application of the concepts of reciprocity (Cowley 1969).

Time-resolved high-resolution electron microscopy of structural stability in mgo clusters

1996 ◽  
Vol 54 ◽  
pp. 672-673
Author(s):  
T. Kizuka ◽  
N. Tanaka
Keyword(s):  
Electron Microscopy ◽  
High Resolution ◽  
Magnesium Oxide ◽  
Structural Stability ◽  
High Resolution Electron Microscopy ◽  
Video Tape ◽  
Solid State Physics ◽  
Time Resolved ◽  
Resolution Electron ◽  
Structural And Functional Properties

Structure and stability of atomic clusters have been studied by time-resolved high-resolution electron microscopy (TRHREM). Typical examples are observations of structural fluctuation in gold (Au) clusters supported on silicon oxide films, graphtized carbon films and magnesium oxide (MgO) films. All the observations have been performed on the clusters consisted of single metal element. Structural stability of ceramics clusters, such as metal-oxide, metal-nitride and metal-carbide clusters, has not been observed by TRHREM although the clusters show anomalous structural and functional properties concerning to solid state physics and materials science.In the present study, the behavior of ceramic, magnesium oxide (MgO) clusters is for the first time observed by TRHREM at 1/60 s time resolution and at atomic resolution down to 0.2 nm.MgO and gold were subsequently deposited on sodium chloride (001) substrates. The specimens, single crystalline MgO films on which Au particles were dispersed were separated in distilled water and observed by using a 200-kV high-resolution electron microscope (JEOL, JEM2010) equipped with a high sensitive TV camera and a video tape recorder system.

Cryo-TEM of amphiphilic polymer and amphiphile/polymer solutions

1993 ◽  
Vol 51 ◽  
pp. 876-877
Author(s):  
Yeshayahu Talmon
Keyword(s):  
Microstructural Characterization ◽  
Building Blocks ◽  
Quantitative Information ◽  
Physical Models ◽  
Specimen Preparation ◽  
Time Resolved ◽  
Temperature Changes ◽  
Temperature And Humidity ◽  
Microstructured Fluids ◽  
Air Streams

To achieve complete microstructural characterization of self-aggregating systems, one needs direct images in addition to quantitative information from non-imaging, e.g., scattering or Theological measurements, techniques. Cryo-TEM enables us to image fluid microstructures at better than one nanometer resolution, with minimal specimen preparation artifacts. Direct images are used to determine the “building blocks” of the fluid microstructure; these are used to build reliable physical models with which quantitative information from techniques such as small-angle x-ray or neutron scattering can be analyzed.To prepare vitrified specimens of microstructured fluids, we have developed the Controlled Environment Vitrification System (CEVS), that enables us to prepare samples under controlled temperature and humidity conditions, thus minimizing microstructural rearrangement due to volatile evaporation or temperature changes. The CEVS may be used to trigger on-the-grid processes to induce formation of new phases, or to study intermediate, transient structures during change of phase (“time-resolved cryo-TEM”). Recently we have developed a new CEVS, where temperature and humidity are controlled by continuous flow of a mixture of humidified and dry air streams.

The promises and perfidy of cryomicroscopy and analysis

1994 ◽  
Vol 52 ◽  
pp. 382-383
Author(s):  
Patrick Echlin
Keyword(s):  
Low Temperature ◽  
High Energy ◽  
Short Paper ◽  
List Type ◽  
Time Resolved ◽  
Energy Beam ◽  
Cellular Analysis ◽  
Dissolved Ions ◽  
Embedding Medium ◽  
Low Temperature Microscopy

The unusual title of this short paper and its accompanying tutorial is deliberate, because the intent is to investigate the effectiveness of low temperature microscopy and analysis as one of the more significant elements of the less interventionist procedures we can use to prepare, examine and analyse hydrated and organic materials in high energy beam instruments. The promises offered by all these procedures are well rehearsed and the litany of petitions and responses may be enunciated in the following mantra.Vitrified water can form the perfect embedding medium for bio-organic samples.Frozen samples provide an important, but not exclusive, milieu for the in situ sub-cellular analysis of the dissolved ions and electrolytes whose activities are central to living processes.The rapid conversion of liquids to solids provides a means of arresting dynamic processes and permits resolution of the time resolved interactions between water and suspended and dissolved materials.The low temperature environment necessary for cryomicroscopy and analysis, diminish, but alas do not prevent, the deleterious side effects of ionizing radiation.Sample contamination is virtually eliminated.

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