Three-dimensional reconstruction of the 30S ribosomal subunit from randomly oriented particles

1983 ◽  
Vol 41 ◽  
pp. 758-759
Author(s):  
A. Verschoor ◽  
J. Frank ◽  
M. Radermacher ◽  
T. Wagenknecht ◽  
M. Boublik
Keyword(s):  
Correspondence Analysis ◽  
Dimensional Space ◽  
Three Dimensional ◽  
Ribosomal Subunit ◽  
Two Dimensional ◽  
Mapping Algorithm ◽  
E Coli ◽  
Wide Range ◽  
Dimensional Reconstruction ◽  
Significant Factors

The small (30S) subunit of prokaryotic ribosomes can assume any of a wide range of tilt positions on the specimen support. Correspondence analysis should make it possible to order views appearing in the electron micrograph according to the angle of tilt.231 individual windowed images from two micrographs showing negatively stained 30S subunits from E. coli ribosomes were subjected to multireference alignment. Correspondence analysis yielded six morphologically significant factors of variance. The second of these related to variations in stain concentrations, which are irrelevant at the level of gross morphology. The coordinates for each image in five-dimensional space (relating to factors 1,3,4,5, and 6) were subjected to a nonlinear mapping algorithm, which calculated an optimal two-dimensional map.The resulting distribution (Fig 1) consisted of two clusters, one of rightfacing, the other of left-facing views. Subaverages along the outer margin of the cluster on the left showed the particle in a range of typical views.

Three-Dimensional Reconstruction of Two Forms of the Chaperonin GRO EL

1990 ◽  
Vol 48 (1) ◽  
pp. 258-259
Author(s):  
J.L. Carrascosa ◽  
G. Abella ◽  
S. Marco ◽  
M. Muyal ◽  
J.M. Carazo
Keyword(s):  
Three Dimensional ◽  
The Other ◽  
Two Dimensional ◽  
Series Method ◽  
Three Dimensional Reconstruction ◽  
Structural Components ◽  
E Coli ◽  
Dimensional Reconstruction ◽  
Morphogenetic Factors ◽  
Tilt Series

Chaperonins are a class of proteins characterized by their role as morphogenetic factors. They trantsiently interact with the structural components of certain biological aggregates (viruses, enzymes etc), promoting their correct folding, assembly and, eventually transport. The groEL factor from E. coli is a conspicuous member of the chaperonins, as it promotes the assembly and morphogenesis of bacterial oligomers and/viral structures.We have studied groEL-like factors from two different bacteria:E. coli and B.subtilis. These factors share common morphological features , showing two different views: one is 6-fold, while the other shows 7 morphological units. There is also a correlation between the presence of a dominant 6-fold view and the fact of both bacteria been grown at low temperature (32°C), while the 7-fold is the main view at higher temperatures (42°C). As the two-dimensional projections of groEL were difficult to interprete, we studied their three-dimensional reconstruction by the random conical tilt series method from negatively stained particles.

Computer Assisted Image Reconstruction of Oligomer Structure

1976 ◽  
Vol 34 ◽  
pp. 472-473
Author(s):  
A.M. Jones ◽  
A. Max Fiskin
Keyword(s):  
Three Dimensional ◽  
Depth Of Field ◽  
Computer Assisted ◽  
Projection Data ◽  
Two Dimensional ◽  
Single Particles ◽  
Dimensional Reconstruction ◽  
Computer Assisted Image ◽  
The Fourier Transform ◽  
Space Approach

If the tilt of a specimen can be varied either by the strategy of observing identical particles orientated randomly or by use of a eucentric goniometer stage, three dimensional reconstruction procedures are available (l). If the specimens, such as small protein aggregates, lack periodicity, direct space methods compete favorably in ease of implementation with reconstruction by the Fourier (transform) space approach (2). Regardless of method, reconstruction is possible because useful specimen thicknesses are always much less than the depth of field in an electron microscope. Thus electron images record the amount of stain in columns of the object normal to the recording plates. For single particles, practical considerations dictate that the specimen be tilted precisely about a single axis. In so doing a reconstructed image is achieved serially from two-dimensional sections which in turn are generated by a series of back-to-front lines of projection data.

Visualisation of E. coli ribosomal RNA in situ by electron spectroscopic imaging and image averaging

1992 ◽  
Vol 50 (1) ◽  
pp. 466-467
Author(s):  
Daniel Beniac ◽  
George Harauz
Keyword(s):  
Ribosomal Rna ◽  
Three Dimensional ◽  
Spectroscopic Imaging ◽  
Electron Spectroscopic Imaging ◽  
E Coli ◽  
Microscopical Technique ◽  
Quantitative Image ◽  
Dimensional Reconstruction ◽  
Image Averaging ◽  
Protein Components

The structures of E. coli ribosomes have been extensively probed by electron microscopy of negatively stained and frozen hydrated preparations. Coupled with quantitative image analysis and three dimensional reconstruction, such approaches are worthwhile in defining size, shape, and quaternary organisation. The important question of how the nucleic acid and protein components are arranged with respect to each other remains difficult to answer, however. A microscopical technique that has been proposed to answer this query is electron spectroscopic imaging (ESI), in which scattered electrons with energy losses characteristic of inner shell ionisations are used to form specific elemental maps. Here, we report the use of image sorting and averaging techniques to determine the extent to which a phosphorus map of isolated ribosomal subunits can define the ribosomal RNA (rRNA) distribution within them.

Polymorphic phase transition of a membrane protein - analysis of continuous diffuse electron diffraction intensity

1986 ◽  
Vol 44 ◽  
pp. 166-167
Author(s):  
Douglas L. Dorset ◽  
Andrew K. Massalski
Keyword(s):  
Molecular Sieve ◽  
Three Dimensional ◽  
Pore Geometry ◽  
Two Dimensional ◽  
Diffraction Intensity ◽  
Polymorphic Phase Transition ◽  
E Coli ◽  
Crystallographic Study ◽  
Hexagonal Form ◽  
Polymorphic Forms

Matrix porin, the ompF gene product of E. coli, has been the object of a electron crystallographic study of its pore geometry in an attempt to understand its function as a membrane molecular sieve. Three polymorphic forms have been found for two-dimensional crystals reconstituted in phospholipid, two hexagonal forms with different lipid content and an orthorhombic form coexisting with and similar to the hexagonal form found after lipid loss. In projection these have been shown to retain the same three-fold pore triplet geometry and analyses of three-dimensional data reveal that the small hexagonal and orthorhombic polymorphs have similar structure as well as unit cell spacings.

Three-dimensional reconstruction of the 40S ribosomal subunit from rabbit reticulocytes

1987 ◽  
Vol 45 ◽  
pp. 936-937
Author(s):  
Neng-Yu Zhang ◽  
Terence Wagenknecht ◽  
Michael Radermacher ◽  
Tom Obrig ◽  
Joachim Frank
Keyword(s):  
Three Dimensional ◽  
Ribosomal Subunit ◽  
Uranyl Acetate ◽  
Reconstruction Method ◽  
Single Exposure ◽  
Electron Dose ◽  
Ribosomal Subunits ◽  
40S Ribosomal Subunit ◽  
Dimensional Reconstruction ◽  
Rabbit Reticulocytes

We have reconstructed the 40S ribosomal subunit at a resolution of 4 nm using the single-exposure pseudo-conical reconstruction method of Radermacher et al.Small (40S) ribosomal subunits were Isolated from rabbit reticulocytes, applied to grids and negatively stained (0.5% uranyl acetate) in a manner that “sandwiches” the specimen between two layers of carbon. Regions of the grid exhibiting uniform and thick staining were identified and photographed twice (magnification 49,000X). The first micrograph was always taken with the specimen tilted by 50° and the second was of the Identical area untilted (Fig. 1). For each of the micrographs the specimen was subjected to an electron dose of 2000-3000 el/nm2.Three hundred thirty particles appearing in the L view (defined in [4]) were selected from both tilted- and untilted-specimen micrographs. The untilted particles were aligned and their rotational alignment produced the azimuthal angles of the tilted particles in the conical tilt series.

Three-Dimensional Structure of Cloned T3 Connector Protein at 1.6nm Resolution

1990 ◽  
Vol 48 (1) ◽  
pp. 282-283
Author(s):  
José L. Carrascosa ◽  
José M. Valpuesta ◽  
Hisao Fujisawa
Keyword(s):  
Dna Sequences ◽  
Expression Vector ◽  
Three Dimensional ◽  
Deae Cellulose ◽  
Dna Packaging ◽  
Dimensional Structure ◽  
Two Dimensional ◽  
Freezing And Thawing ◽  
Three Dimensional Structure ◽  
Dimensional Reconstruction

The head to tail connector of bacteriophages plays a fundamental role in the assembly of viral heads and DNA packaging. In spite of the absence of sequence homology, the structure of connectors from different viruses (T4, Ø29, T3, P22, etc) share common morphological features, that are most clearly revealed in their three-dimensional structure. We have studied the three-dimensional reconstruction of the connector protein from phage T3 (gp 8) from tilted view of two dimensional crystals obtained from this protein after cloning and purification.DNA sequences including gene 8 from phage T3 were cloned, into Bam Hl-Eco Rl sites down stream of lambda promotor PL, in the expression vector pNT45 under the control of cI857. E R204 (pNT89) cells were incubated at 42°C for 2h, harvested and resuspended in 20 mM Tris HC1 (pH 7.4), 7mM 2 mercaptoethanol, ImM EDTA. The cells were lysed by freezing and thawing in the presence of lysozyme (lmg/ml) and ligthly sonicated. The low speed supernatant was precipitated by ammonium sulfate (60% saturated) and dissolved in the original buffer to be subjected to gel nitration through Sepharose 6B, followed by phosphocellulose colum (Pll) and DEAE cellulose colum (DE52). Purified gp8 appeared at 0.3M NaCl and formed crystals when its concentration increased above 1.5 mg/ml.

scholarly journals Three-Dimensional Thematic Map Imaging of the Yacht Port on the Example of the Polish National Sailing Centre Marina in Gdańsk

2021 ◽  
Vol 11 (15) ◽  
pp. 7016
Author(s):  
Pawel S. Dabrowski ◽  
Cezary Specht ◽  
Mariusz Specht ◽  
Artur Makar
Keyword(s):  
Spatial Data ◽  
Convex Surface ◽  
Dimensional Space ◽  
Three Dimensional ◽  
Future Research ◽  
Two Dimensional ◽  
Thematic Maps ◽  
Research Directions ◽  
Future Research Directions ◽  
The Many

The theory of cartographic projections is a tool which can present the convex surface of the Earth on the plane. Of the many types of maps, thematic maps perform an important function due to the wide possibilities of adapting their content to current needs. The limitation of classic maps is their two-dimensional nature. In the era of rapidly growing methods of mass acquisition of spatial data, the use of flat images is often not enough to reveal the level of complexity of certain objects. In this case, it is necessary to use visualization in three-dimensional space. The motivation to conduct the study was the use of cartographic projections methods, spatial transformations, and the possibilities offered by thematic maps to create thematic three-dimensional map imaging (T3DMI). The authors presented a practical verification of the adopted methodology to create a T3DMI visualization of the marina of the National Sailing Centre of the Gdańsk University of Physical Education and Sport (Poland). The profiled characteristics of the object were used to emphasize the key elements of its function. The results confirmed the increase in the interpretative capabilities of the T3DMI method, relative to classic two-dimensional maps. Additionally, the study suggested future research directions of the presented solution.

Symbol recognition system based on 3D stereo vision

2020 ◽  
pp. 1-10
Author(s):  
Linlin Wang
Keyword(s):  
Camera Calibration ◽  
Stereo Vision ◽  
Stereo Matching ◽  
Dimensional Space ◽  
Three Dimensional ◽  
Recognition System ◽  
Symbol Recognition ◽  
Two Dimensional ◽  
Measuring Machine ◽  
Left And Right

With the continuous development of computer science and technology, symbol recognition systems may be converted from two-dimensional space to three-dimensional space. Therefore, this article mainly introduces the symbol recognition system based on 3D stereo vision. The three-dimensional image is taken by the visual coordinate measuring machine in two places on the left and right. Perform binocular stereo matching on the edge of the feature points of the two images. A corner detection algorithm combining SUSAN and Harris is used to detect the left and right camera calibration templates. The two-dimensional coordinate points of the object are determined by the image stereo matching module, and the three-dimensional discrete coordinate points of the object space can be obtained according to the transformation relationship between the image coordinates and the actual object coordinates. Then draw the three-dimensional model of the object through the three-dimensional drawing software. Experimental data shows that the logic resources and memory resources occupied by image preprocessing account for 30.4% and 27.4% of the entire system, respectively. The results show that the system can calibrate the internal and external parameters of the camera. In this way, the camera calibration result will be more accurate and the range will be wider. At the same time, it can effectively make up for the shortcomings of traditional modeling techniques to ensure the measurement accuracy of the detection system.

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