The Structure of the Macromolecular Units on the Cell Walls of Bacillus Polymyxa

1967 ◽  
Vol 2 (4) ◽  
pp. 587-591
Author(s):  
J. T. FINCH ◽  
A. KLUG ◽  
M.V. NERMUT

Electron micrographs of negatively stained preparations of cell walls of Bacillus polymyxa have been investigated by optical diffraction and optical filtering techniques. Images of single layers of the cell wall, from which the ‘noise’ has been filtered optically, show hollow, square-shaped morphological units arranged on a square lattice of side 100 Å. Single-layer images showing the same pattern have been filtered from moiré patterns arising from two overlapping single layers. The morphological units are composed of four smaller subunits. The optical diffraction patterns from regions of two overlapping layers show extra reflexions which are attributed to multiple electron scattering.

Author(s):  
Murray Stewart ◽  
T.J. Beveridge ◽  
D. Sprott

The archaebacterium Methanospirillum hungatii has a sheath as part of its cell wall which is composed mainly of protein. Treatment with dithiothreitol or NaOH released the intact sheaths and electron micrographs of this material negatively stained with uranyl acetate showed flattened hollow tubes, about 0.5 μm diameter and several microns long, in which the patterns from the top and bottom were superimposed. Single layers, derived from broken tubes, were also seen and were more simply analysed. Figure 1 shows the general appearance of a single layer. There was a faint axial periodicity at 28.5 A, which was stronger at irregular multiples of 28.5 A (3 and 4 times were most common), and fine striations were also seen at about 3° to the tube axis. Low angle electron diffraction patterns (not shown) and optical diffraction patterns (Fig. 2) from these layers showed a complex meridian (as a result of the irregular nature of the repeat along the tube axis) which showed a clear maximum at 28.5 A, consistent with the basic subunit spacing.


We have used the techniques of optical diffraction and optical filtering to study electron micrographs of myofibrils and of paracrystals of myofibrillar proteins. The optical diffraction patterns provide information about periodic structure in the micrographs, and sometimes may reveal periodicities not apparent to the eye. We compare the optical diffraction patterns with the X-ray diffraction patterns obtained from living muscle, and this comparison can assist our interpretation of both the X-ray diffraction patterns and the electron micrographs. The optical diffractometer we have used is essentially similar to those described by Taylor & Lipson (1964), and by Klug & DeRosier (1966). The apparatus incorporates several refinements to facilitate operation. The recombining lens has a focal length, f , of about 1 m, and is placed so that the recombined image is formed at 2 f and has the same size as the subject. The diffraction subjects are not usually the electron micrographs themselves but copies on film. The film is of more uniform optical thickness than the glass electron micrograph, and is less fragile. Moreover, a set of films of varying contrast can be made from one micrograph.


1977 ◽  
Vol 75 (3) ◽  
pp. 818-836 ◽  
Author(s):  
M A Goldstein ◽  
J P Schroeter ◽  
R L Sass

Optical diffraction patterns from electron micrographs of both longitudinal and cross sections of normal and anomalous canine cardiac Z bands have been compared. The data indicate that anomalous cardiac Z bands resembling nemaline rods are structurally related to Z bands in showing a repeating lattice common to both. In thin sections transverse to the myofibril axis, both electron micrographs and optical diffraction patterns of the Z structure reveal a square lattice of 24 nm. This lattice is simple at the edge of each I band and centered in the interior of the Z band, where two distinct lattice forms have been observed. In longitudinal sections, oblique filaments visible in the electron micrographs correspond to a 38-nm axial periodicity in diffraction patterns of both Z band and Z rod. We conclude that the Z rods will be useful for further analysis and reconstruction of the Z lattice by optical diffraction techniques.


Author(s):  
Glen B. Haydon

Analysis of light optical diffraction patterns produced by electron micrographs can easily lead to much nonsense. Such diffraction patterns are referred to as optical transforms and are compared with transforms produced by a variety of mathematical manipulations. In the use of light optical diffraction patterns to study periodicities in macromolecular ultrastructures, a number of potential pitfalls have been rediscovered. The limitations apply to the formation of the electron micrograph as well as its analysis.(1) The high resolution electron micrograph is itself a complex diffraction pattern resulting from the specimen, its stain, and its supporting substrate. Cowley and Moodie (Proc. Phys. Soc. B, LXX 497, 1957) demonstrated changing image patterns with changes in focus. Similar defocus images have been subjected to further light optical diffraction analysis.


1970 ◽  
Vol 16 (9) ◽  
pp. 883-887 ◽  
Author(s):  
R. G. E. Murray ◽  
Myrtle M. Hall ◽  
J. Marak

Sections of germinating spores of Bacillus polymyxa show that the primordial cell wall consists of a single layer. The intermediate layer and an outer rectangular array of macromolecules found on vegetative cells do not appear until the spore coats crack open about 60 min after initiation of germination. The initial areas of the new components appear in patches under the cracks in the coats. Within 10 min the wall is completed and takes on the profile seen in the vegetative cell. Negative staining and freeze-etching techniques show the regular structure to be identical with that previously shown for mature cells, although the subunits are more readily visible in negatively stained preparations.


1966 ◽  
Vol 12 (1) ◽  
pp. 105-108 ◽  
Author(s):  
K. Jane Carson ◽  
R. G. Eagon

Electron micrographs of thin sections of normal cells of Pseudomonas aeruginosa showed the cell walls to be convoluted and to be composed of two distinct layers. Electron micrographs of thin sections of lysozyme-treated cells of P. aeruginosa showed (a) that the cell walls lost much of their convoluted nature; (b) that the layers of the cell walls became diffuse and less distinct; and (c) that the cell walls became separated from the protoplasts over extensive cellular areas. These results suggest that the peptidoglycan component of the unaltered cell walls of P. aeruginosa is sensitive to lysozyme. Furthermore, it appears that the peptidoglycan component is not solely responsible for the rigidity of the cell walls of Gram-negative bacteria.


1975 ◽  
Vol 99 (1) ◽  
pp. 143-151 ◽  
Author(s):  
Douglas H. Ohlendorf ◽  
Myra L. Collins ◽  
Leonard J. Banaszak

1982 ◽  
Vol 92 (2) ◽  
pp. 443-451 ◽  
Author(s):  
R W Kensler ◽  
R J Levine

Long, thick filaments (greater than 4.0 micrometer) rapidly and gently isolated from fresh, unstimulated Limulus muscle by an improved procedure have been examined by electron microscopy and optical diffraction. Images of negatively stained filaments appear highly periodic with a well-preserved myosin cross-bridge array. Optical diffraction patterns of the electron micrographs show a wealth of detail and are consistent with a myosin helical repeat of 43.8 nm, similar to that observed by x-ray diffraction. Analysis of the optical diffraction patterns, in conjunction with the appearance in electron micrographs of the filaments, supports a model for the filament in which the myosin cross-bridges are arranged on a four-stranded helix, with 12 cross-bridges per turn or each helix, thus giving an axial repeat every third level of cross-bridges (43.8 nm).


Author(s):  
Gonpachiro Yasuzumi

The fine structure of the crystalline intranuclear inclusions in the human osteosarcoma cells was studied by using a goniometer which tilted the specimen at angles of ±30° and ±40°. The results appear in the series of micrographs showing Fig. 1. At the point by the arrow, a helical structure is visible in two filaments. Optical diffrection patterns of selected areas of each negative electron micrograph film were taken by using a helium-neon laser as a source.A tentative model of the structure can be based on optical diffraction techniques applied to electron micrographs taken at different tilt angles. The diffraction pattern taken from image No. 0 in Fig. 1 is depicted in Fig. 2. Diffraction patterns from Nos. 3, 4 and 8 are shown in Figs. 3, 4 and 5. The contrast of some periodic structures can be eliminated in the image (extinction effect due to Bragg reflection of electron waves) by the interference of scattered waves from constituent elements. Hence it is rather important to interpret the electron micrographs and their optical diffraction patterns by considering the extinction effect of the image contrast.


1979 ◽  
Vol 83 (1) ◽  
pp. 187-204 ◽  
Author(s):  
M A Goldstein ◽  
J P Schroeter ◽  
R L Sass

Filtered images of mammalian cardiac Z bands were reconstructed from optical diffraction patterns from electron micrographs. Reconstructed images from longitudinal sections show connecting filaments at each 38-nm axial repeat in an array consistent with cross-sectional data. Some reconstructed images from cross sections indicate two distinctly different optical diffraction patterns, one for each of two lattice forms (basket weave and small square). Other images are more complex and exhibit composite diffraction patterns. Thus, the two lattice forms co-exist, interconvert, or represent two different aspects of the same details within the lattice. Two three-dimensional models of the Z lattice are presented. Both include the following features: a double array of axial filaments spaced at 24 nm, successive layers of tetragonally arrayed connecting filaments, projected fourfold symmetry in cross section, and layers of connecting filaments spaced at intervals of 38 nm along the myofibril axis. Projected views of the models are compared to electron micrographs and optically reconstructed images of the Z lattice in successively thicker cross sections. The entire Z band is rarely a uniform lattice regardless of plane of section or section thickness. Optical reconstructions strongly suggest two types of variation in the lattice substructure: (a) in the arrangement of connecting filaments, and (b) in the arrangement of units added side-to-side to make larger myofilament bundles and/or end-to-end to make wider Z bands. We conclude that the regular arrangement of axial and connecting filaments generates a dynamic Z lattice.


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