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M Stewart

Publications and source records attributed to M Stewart.

At least 469 records · Page 26Linked to original sources

Interaction of aldolase with actin-containing filaments. Structural studies.

Electron micrographs of the paracrystals formed when fructose bisphosphate aldolase (EC 4.1.2.13) is added to actin-containing filaments were analysed by computer methods so that ultrastructural changes could be correlated with the various stoicheiometries of binding determined in the preceding paper [Walsh, Winzor, Clarke, Masters & Morton (1980) Biochem. J. 186, 89-98]. Paracrystals formed with aldolase and either F-actin or F-actin-tropomyosin have a single light transverse band every 38 nm, which is due to aldolase molecules cross-linking the filaments. In contrast, the paracrystals formed between aldolase and F-actin-tropomyosin-troponin filaments show two transverse bands every 38 nm: a major band, interpreted as aldolase binding to troponin, and a minor band, interpreted as aldolase cross-linking the filaments. The intensity of the minor band varies with Ca2+ concentration, being greatest when the Ca2+ concentration is low. A model for the different paracrystal structures which relates the various patterns and binding stoicheiometries to structural changes in the actin-containing filaments is proposed.

Actins↗

Filament formation in smooth muscle homogenates.

To provide more detailed information on the aggregation properties of smooth muscle myosin, we have extended earlier work on the formation of thick filaments when homogenates of guinea-pig taenia coli and chicken gizzard muscle are diluted. In both preparations there is a slow and a fast phase of filament formation. The slow phase, which generally develops over several hours, appears to depend primarily on the ATP concentration while the rapid phase, which develops over 5-15 min, is influenced by the extent of dilution, homogenization conditions, divalent cation concentration, ATP concentration and presence of chicken gizzard tropomyosin. Many of these effects on the rapid phase can be explained by postulating that filament formation only takes place when the ATP concentration is reduced. There are significant differences between the filament populations formed from each muscle, with those from taenia coli being shorter than those from gizzard. Two types of filament are present in preparations from each muscle, the first being characterized by the presence of a central bare zone and cross striations at both ends, whilst the second have cross striations along their entire length; the periodicity of the cross striations appears to be 14.5 nm. The bare zone filaments have an average length and width of 325 nm and 17.6 nm respectively, while the corresponding values for the cross striated filaments is 3 : 1 for taenia coli and 1 : 3 for chicken gizzard, which accounts for the difference in average filament length observed between these preparations. The gizzard filaments appear to form more readily than those of taenia coli.

Animals↗

Structure of the regular surface layer of Sporosarcina ureae.

Optical diffraction and computer image processing of electron micrographs were employed to analyze the structure of the regular surface layer of Sporosarcina ureae at high resolution. Negatively stained preparations of regular surface layer fragments showed two types of tetragonal pattern, each having p4 symmetry in projection with a = 12.8 nm. Although the two patterns differed greatly in overall appearance, both had a common pattern of areas of high stain density which we interpret as arising from gaps or holes in the structure. We speculate that these holes may be related to a protective role of the regular surface layer, whereby hostile environmental agents (such as muramidases) larger than about 2 nm would be screened from the underlying layers of the bacterial surface, while the free passage of nutrients and waste products into and out of the cell would still be allowed.

Bacillaceae↗

Distribution of calcium and other elements in cryosectioned Bacillus cereus T spores, determined by high-resolution scanning electron probe x-ray microanalysis.

The distribution of a number of key elements in Bacillus cereus T spores was determined by high-resolution scanning electron probe X-ray microanalysis. To circumvent the redistribution of soluble or weakly bound elements, freeze-dried cryosections of spores, which had been rapidly frozen in 50% aqueous polyvinyl pyrrolidone, were employed. The sections were examined by using a modified Philips EM400 electron microscope fitted with a field emission gun, scanning transmission electron microscopy attachment, and a computer-linked energy-dispersive X-ray microanalysis system. X-ray maps for selected elements and the corresponding electron image were produced simultaneously by scanning the cryosections with a fine electron beam in a raster pattern, using the scanning transmission electron microscopy attachment. The results indicated that almost all of the calcium, magnesium, and manganese, together with most of the phosphorus, was located in the core region. An unexpectedly high concentration of silicon was found in the cortex/coat layer. Granules containing high concentrations of calcium, manganese, and phosphorus were demonstrated in spores containing reduced levels of dipicolinic acid. Spot mode analyses, in which a stationary beam was located over the region of interest in the spore cryosection, confirmed the results obtained with the scanning mode and also provided a more accurate quantitation of the elemental concentrations on a dry weight bases.

Bacillus cereus↗

Changes associated with glycolytic-enzyme binding in the equatorial X-ray-diffraction pattern of glycerinated rabbit psoas muscle.

The binding of fructose biphosphate aldolase to the thin filaments of glycerinated rabbit psoas muscle produces a significant change in its low-angle X-ray-diffraction pattern. The intensity of the (11) reflection relative to that of the (10) reflection increases by 26 +/- 3% (mean +/- S.E.M.), which is consistent with the increase in the mass of the thin filaments produced by enzyme binding. A similar effect is found with a mixture of aldolase and glyceraldehyde 3-phosphate dehydrogenase. The significance of the change in intensity is considered with reference to the interpretation of the equatorial patterns obtained from muscles in different physiological states. The magnitude of the increase in the relative intensity of the (11) reflection is lower than that observed between relaxed and contracting muscle and does not bring into question the interpretation linking changes in these patterns to cross-bridge movement. However, the effect due to enzyme binding may be important when making detailed interpretations of these changes. It may also be related to an unusual pattern sometimes observed in cardiac muscle.

Animals↗

An electron-microscope study of beta-glucuronidase crystals.

beta-Glucuronidase from rat preputial glands was crystallized as thin sheets having p6 symmetry in projection with a equal 20.2nm. A filtered image was produced by Fourier methods to a resolution of 2.2 nm by averaging information from six areas. This suggests an approximately triangular molecular outline in projection, and this is taken to indicate a probable tetrahedral arrangement of the four subunits of the beta-glucuronidase molecule.

Computers↗

Hypofibrinogenemia-dysfibrinogenemia and von Willebrand's disease in the same family.

Two Puerto Rican families were studied. One family included a number of members with dysfibrinogenemia occasionally associated with hypofibrinogenemia. The second family had members with von Willebrand's disease. The two diseases merged in the proband's immediate family; the affected members of this family exhibited a mild bleeding disorder. Others in the two families had no obvious bleeding tendency.

Adult↗

Electron probe analysis of muscle and X-ray mapping of biological specimens with a field emission gun.

Recent electron probe analytic studies of freeze-dried cryosections of vascular smooth and vertebrate striated muscle are reviewed. The results show that the sarcoplasmic reticulum of striated muscle is not in ionic communication with the extracellular space. Vacuolation by hypertonic solutions and fatigue involves the T-tubule system. The high calcium content of the terminal cisternae of the resting muscle has been quantitated in situ. In smooth muscle, the high Cl content is distributed in the cytoplasm, and mitochondria in rabbit portal vein smooth muscle cells do not contain high concentrations of calcium. Mitochondrial calcium loading in the form of granules is generally due to fiber damage. Nuclear and mitochondrial composition in situ has been quantitated and compared to the composition of the cytoplasm of the same cells. Preliminary phosphorus x-ray maps of smooth muscle show the feasibility of this approach in defining the composition of organelles in thin cryosections. The use of x-ray maps at intermediate resolution is illustrated with tropomyosin paracrystals labelled with Hg-containing dye at the thiol residues. Mercury x-ray maps of such paracrystals show the 40nm periodicity of the thiol groups and their Fourier transforms contain information to a spatial resolution of 10-20nm.

Animals↗

Heavy meromyosin labeling of intermediate filaments in cultured connective tissue cells.

Mild treatment with trypsin causes a radical change in the heavy meromyosin (HMM) binding properties of intermediate filaments in glycerinated, myosin-extracted cultured chick embryo connective tissue cells. In non-trypsin-treated cells, HMM labeling of filaments was often indistinct and variable in its distribution. By contrast, in cells treated with trypsin (under conditions which allowed most intermediate filaments to survive), virtually all filaments, including those of intermediate size, decorated with HMM to give distinct arrowhead patterns. We suggest that most intermediate filaments in such cells contain a core of F-actin masked by trypsin-labile accessory proteins.

Actins↗

A new look at the assessment of mental development in young children recovering from severe malnutrition.

Previous studies of malnourished children in hospital have shown that their developmental quotients (DQS) improve as their nutritional status improves, but they have not taken into account the possible effects of being in hospital and of test practice. The present study assessed development of malnourished and adequately nourished children in hospital and found that mean DQS of each group rose to a similar extent during recovery from illness. It is concluded that DQS on admission to hospital were lowered by the children's discomfort and unhappiness at being in hospital, and/or by illness, and that rises in DQ shown by children recovering from malnutrition are not necessarily attributable to improving nutritional status.

Child Development↗