scholarly journals Automated electron microscopy for evaluating two-dimensional crystallization of membrane proteins

2010 ◽  
Vol 171 (1) ◽  
pp. 102-110 ◽  
Author(s):  
Minghui Hu ◽  
Martin Vink ◽  
Changki Kim ◽  
KD Derr ◽  
John Koss ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Membrane Proteins ◽  
Two Dimensional

Reconstruction of Two-Dimensional Projections of GP 32*I

1981 ◽  
Vol 39 ◽  
pp. 38-39
Author(s):  
H.A. Cohen ◽  
W. Chiu ◽  
J. Hosoda
Keyword(s):  
Electron Microscopy ◽  
Molecular Weight ◽  
Uranyl Acetate ◽  
Distilled Water ◽  
Acetate Solution ◽  
Two Dimensional ◽  
Parent Molecule ◽  
T4 Bacteriophage ◽  
Electron Imaging ◽  
Better Than

GP 32 (molecular weight 35000) is a T4 bacteriophage protein that destabilizes the DNA helix. The fragment GP32*I (77% of the total weight), which destabilizes helices better than does the parent molecule, crystallizes as platelets thin enough for electron diffraction and electron imaging. In this paper we discuss the structure of this protein as revealed in images reconstructed from stained and unstained crystals.Crystals were prepared as previously described. Crystals for electron microscopy were pelleted from the buffer suspension, washed in distilled water, and resuspended in 1% glucose. Two lambda droplets were placed on grids over freshly evaporated carbon, allowed to sit for five minutes, and then were drained. Stained crystals were prepared the same way, except that prior to draining the droplet, two lambda of aqueous 1% uranyl acetate solution were applied for 20 seconds. Micrographs were produced using less than 2 e/Å2 for unstained crystals or less than 8 e/Å2 for stained crystals.

Parabolas from Reconstructed Surfaces Observed in Convergent-Beam RHEED Patterns

1990 ◽  
Vol 48 (2) ◽  
pp. 398-399
Author(s):  
M. Gajdardziska-Josifovska
Keyword(s):  
Electron Microscopy ◽  
High Energy ◽  
Two Dimensional ◽  
High Energy Electron ◽  
Reflection And Transmission ◽  
Experimental Diffraction Pattern ◽  
Surface Resonance ◽  
Reconstructed Surfaces ◽  
Reflection Electron Microscopy ◽  
Convergent Beam

Parabolas have been observed in the reflection high-energy electron diffraction (RHEED) patterns from surfaces of single crystals since the early thirties. In the last decade there has been a revival of attempts to elucidate the origin of these surface parabolas. The renewed interest stems from the need to understand the connection between the parabolas and the surface resonance (channeling) condition, the latter being routinely used to obtain higher intensity in reflection electron microscopy (REM) images of surfaces. Several rather diverging descriptions have been proposed to explain the parabolas in the reflection and transmission Kikuchi patterns. Recently we have developed an unifying general treatment in which the parabolas are shown to be K-lines of two-dimensional lattices. Here we want to review the main features of this description and present an experimental diffraction pattern from a 30° MgO (111) surface which displays parabolas that can be attributed to the surface reconstruction.

Two-Dimensional Crystallization of Tetanus Toxin

1985 ◽  
Vol 43 ◽  
pp. 528-529
Author(s):  
Jeffry A. Reidler ◽  
John P. Robinson
Keyword(s):  
Electron Microscopy ◽  
Tetanus Toxin ◽  
Structural Information ◽  
Three Dimensional ◽  
Two Dimensional ◽  
Membrane Interface ◽  
3D Reconstructions ◽  
Cellular Receptors ◽  
Computer Reconstruction ◽  
2D Crystals

We have prepared two-dimensional (2D) crystals of tetanus toxin using procedures developed by Uzgiris and Kornberg for the directed production of 2D crystals of monoclonal antibodies at an antigen-phospholipid monolayer interface. The tetanus toxin crystals were formed using a small mole fraction of the natural receptor, GT1, incorporated into phosphatidyl choline monolayers. The crystals formed at low concentration overnight. Two dimensional crystals of this type are particularly useful for structure determination using electron microscopy and computer image refinement. Three dimensional (3D) structural information can be derived from these crystals by computer reconstruction of photographs of toxin crystals taken at different tilt angles. Such 3D reconstructions may help elucidate the mechanism of entry of the enzymatic subunit of toxins into cells, particularly since these crystals form directly on a membrane interface at similar concentrations of ganglioside GT1 to the natural cellular receptors.

Characterization of microvillar membrane proteins of dog small intestine by two-dimensional electrophoresis

1995 ◽  
Vol 110 (3) ◽  
pp. 483-492 ◽  
Author(s):  
Philip W. Pemberton ◽  
Robert W. Lobley ◽  
Raymond Holmes ◽  
Susanne H. Sørensen ◽  
Kenneth W. Simpson ◽  
...  
Keyword(s):  
Small Intestine ◽  
Membrane Proteins ◽  
Two Dimensional ◽  
Two Dimensional Electrophoresis ◽  
Dimensional Electrophoresis
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