scholarly journals MITORIBOSOMES FROM CANDIDA UTILIS

1972 ◽  
Vol 54 (3) ◽  
pp. 468-492 ◽  
Author(s):  
Pierre V. Vignais ◽  
Barbara J. Stevens ◽  
Janine Huet ◽  
Jean André
Keyword(s):  
Candida Utilis ◽  
Buoyant Density ◽  
Electron Microscopic Observation ◽  
Electron Microscopic ◽  
Mitochondrial Ribosomes ◽  
Characteristic Views ◽  
Polypeptide Chains ◽  
Complete Dissociation ◽  
Cytoplasmic Ribosomes ◽  
Made In

Highly purified mitochondrial ribosomes (mitoribosomes) have been obtained from the yeast Candida utilis. Sedimentation analysis in sucrose gradients made in 5 mM MgCl2, 1 mM Tris, pH 7.4 and 50 mM KCl clearly distinguishes mitoribosomes (72S) from cytoplasmic ribosomes (cytoribosomes) (78S). Mitoribosomes are completely dissociated into 50S and 36S subunits at 10-4 M MgCl2 whereas complete dissociation of cytoribosomes into 61S and 37S subunits occurs only at 10-6 M MgCl2 Electron microscopy of negatively stained mitoribosomes (72S peak) shows bipartite profiles, about 265 x 210 x 200 A Characteristic views are interpreted as frontal, dorsal, and lateral projections of the particles, the latter is observed in two enantiomorphic forms Mitoribosome 50S subunits display rounded profiles bearing a conspicuous knoblike projection, reminiscent of the large bacterial subunit. The 36S subunits show a variety of angular profiles. Mitoribosomal subunits are subject to artifactual dimerization at high Mg2+ concentration Under these conditions, a supplementary 80S peak arises. Electron microscopic observation of the 80S peak reveals closely paired particles of the 50S type Buoyant density determinations after glutaraldehyde fixation show a single peak at ρ = 1.48 for mitoribosomes and 1.53 for cytoribosomes In the presence of ethylenediaminetetraacetate (EDTA), two species of RNA, 21S and 16S, are obtained from mitoribosomes, while 25S and 17S RNA are obtained from cytoribosomes It is established that the small and large RNA species are derived from the 36S and 50S subunits, respectively, by extraction of the RNA from each subunit The G + C content of the RNA is lower for mitoribosomes (33%) than for cytoribosomes (50%). Incubation of C utilis mitochondria with leucine-14C results in the labeling of 72S mitoribosomes. The leucine-14C incorporation is inhibited by chloramphenicol and resistant to cycloheximide Puromycin strips the incorporated radioactivity from the 72S mitoribosomes, which is consistent with the view that leucine-14C is incorporated into nascent polypeptide chains at the level of mitoribosomes

scholarly journals Mitochondrial and cytoplasmic ribosomes from mammalian tissues. Further characterization of ribosomal subunits and validity of buoyant-density methods for determination of the chemical composition and partial specific volume of ribonucleoprotein particles

1977 ◽  
Vol 168 (2) ◽  
pp. 245-259 ◽  
Author(s):  
Ada Sacchi ◽  
Ugo Ferrini ◽  
Paola Londei ◽  
Piero Cammarano ◽  
Nadir Maraldi
Keyword(s):  
Chemical Composition ◽  
Specific Volume ◽  
Buoyant Density ◽  
Volume Ratio ◽  
Partial Specific Volume ◽  
Ribosomal Subunits ◽  
Molecular Weights ◽  
Mitochondrial Ribosomes ◽  
Density Methods ◽  
Cytoplasmic Ribosomes

1. At 0–4°C mitochondrial ribosomes (55S) dissociate into 39S and 29S subunits after exposure to 300mm-K+ in the presence of 3.0mm-Mg2+. When these subunits are placed in a medium containing a lower concentration of K+ ions (25mm), approx. 75% of the subparticles recombine giving 55S monomers. 2. After negative staining the large subunits (20.3nm width) usually show a roundish profile, whereas the small subunits (12nm width) show an elongated, often bipartite, profile. The dimensions of the 55S ribosomes are 25.5nm×20.0nm×21.0nm, indicating a volume ratio of mitochondrial to cytosol ribosomes of 1:1.5. 3. The 39S and 29S subunits obtained in high-salt media at 0–4°C have a buoyant density of 1.45g/cm3; from the rRNA content calculated from buoyant density and from the rRNA molecular weights it is confirmed that the two subparticles have weights of 2.0×106 daltons and 1.20×106 daltons; the weights of the two subunits of cytosol ribosomes are 2.67×106 and 1.30×106 daltons. 4. The validity of the isodensity-equilibrium-centrifugation methods used to calculate the chemical composition of ribosomes was reinvestigated; it is confirmed that (a) reaction of ribosomal subunits with 6.0% (v/v) formaldehyde at 0°C is sufficient to fix the particles, so that they remain essentially stable after exposure to dodecyl sulphate or centrifugation in CsCl, and (b) the partial specific volume of ribosomal subunits is a simple additive function of the partial specific volumes of RNA and protein. The RNA content is linearly related to buoyant density by the equation RNA (% by wt.)=349.5−(471.2×1/ρCsCl), where 1/ρCsCl=[unk]RNP (partial specific volume of ribonucleoprotein). 5. The nucleotide compositions of the two subunit rRNA species of mitochondrial ribosomes from rodents (42% and 43% G+C) are distinctly different from those of cytoplasmic ribosomes.

Properties of Isolated Mitochondria of Agaricus Bisporus: Their Relationship to Dormancy and the Germination of Spores

1973 ◽  
Vol 31 ◽  
pp. 458-459
Author(s):  
K. S. McCarty ◽  
R. F. Weave ◽  
L. Kemper ◽  
F. S. Vogel
Keyword(s):  
Mitochondrial Dna ◽  
Ethidium Bromide ◽  
Microscopic Examination ◽  
Agaricus Bisporus ◽  
Osmotic Shock ◽  
Electron Microscopic Examination ◽  
Electron Microscopic ◽  
Isolated Mitochondria ◽  
Dna Strands ◽  
Cytoplasmic Ribosomes

During the prodromal stages of sporulation in the Basidiomycete, Agaricus bisporus, mitochondria accumulate in the basidial cells, zygotes, in the gill tissues prior to entry of these mitochondria, together with two haploid nuclei and cytoplasmic ribosomes, into the exospores. The mitochondria contain prominent loci of DNA [Fig. 1]. A modified Kleinschmidt spread technique1 has been used to evaluate the DNA strands from purified whole mitochondria released by osmotic shock, mitochondrial DNA purified on CsCl gradients [density = 1.698 gms/cc], and DNA purified on ethidium bromide CsCl gradients. The DNA appeared as linear strands up to 25 u in length and circular forms 2.2-5.2 u in circumference. In specimens prepared by osmotic shock, many strands of DNA are apparently attached to membrane fragments [Fig. 2]. When mitochondria were ruptured in hypotonic sucrose and then fixed in glutaraldehyde, the ribosomes were released for electron microscopic examination.

Electron Microscopic Observation of the Longitudinal Organization of Poly (p-Phenylene Terephthalamide) Fibers

1982 ◽  
Vol 40 ◽  
pp. 680-681
Author(s):  
Li Li-Sheng ◽  
L.F. Allard ◽  
W.C. Bigelow
Keyword(s):  
Electron Microscopy ◽  
Mechanical Properties ◽  
Scanning Electron Microscopy ◽  
Microscopic Observation ◽  
Electron Microscopic Observation ◽  
Electron Microscopic ◽  
Aromatic Polyamides ◽  
Radial Arrangement ◽  
Scanning Electron ◽  
Better Than

The aromatic polyamides form a class of fibers having mechanical properties which are much better than those of aliphatic polyamides. Currently, the accepted morphology of these fibers as proposed by M.G. Dobb, et al. is a radial arrangement of pleated sheets, with the plane of the pleats parallel to the axis of the fiber. We have recently obtained evidence which supports a different morphology of this type of fiber, using ultramicrotomy and ion-thinning techniques to prepare specimens for transmission and scanning electron microscopy.

Observation of Concanavalin-A receptors on hydrated planar erythrocyte membrane film by in situ electron microscopy

1983 ◽  
Vol 41 ◽  
pp. 608-609
Author(s):  
Neng-Bo He ◽  
S.W. Hui
Keyword(s):  
Lipid Bilayers ◽  
Erythrocyte Membrane ◽  
Lipid Membranes ◽  
Surface Film ◽  
Biological Membranes ◽  
Electron Microscopic Observation ◽  
Electron Microscopic ◽  
Black Lipid Membranes ◽  
Fine Mesh ◽  
Planar Films

Monolayers and planar "black" lipid membranes have been widely used as models for studying the structure and properties of biological membranes. Because of the lack of a suitable method to prepare these membranes for electron microscopic observation, their ultrastructure is so far not well understood. A method of forming molecular bilayers over the holes of fine mesh grids was developed by Hui et al. to study hydrated and unsupported lipid bilayers by electron diffraction, and to image phase separated domains by diffraction contrast. We now adapted the method of Pattus et al. of spreading biological membranes vesicles on the air-water interfaces to reconstitute biological membranes into unsupported planar films for electron microscopic study. hemoglobin-free human erythrocyte membrane stroma was prepared by hemolysis. The membranes were spreaded at 20°C on balanced salt solution in a Langmuir trough until a surface pressure of 20 dyne/cm was reached. The surface film was repeatedly washed by passing to adjacent troughs over shallow partitions (fig. 1).

The Scanning Electron Microscopic Observation of the Vestibular Sensory Epithelia

1969 ◽  
Vol 27 ◽  
pp. 40-41
Author(s):  
D.J. Lim ◽  
W.C. Lane
Keyword(s):  
Semicircular Canals ◽  
Electron Microscopic Observation ◽  
Gravitational Acceleration ◽  
Electron Microscopic ◽  
Sensory Epithelia ◽  
Scanning Electron Microscopic ◽  
Vestibular Sensory Epithelia ◽  
And Function ◽  
Sand Piles ◽  
Active Investigation

The morphology and function of the vestibular sensory organs has been extensively studied during the last decade with the advent of electron microscopy and electrophysiology. The opening of the space age also accelerated active investigation in this area, since this organ is responsible for the sensation of balance and of linear, angular and gravitational acceleration.The vestibular sense organs are formed by the saccule, utricle and three ampullae of the semicircular canals. The maculae (sacculi and utriculi) have otolithic membranes on the top of the sensory epithelia. The otolithic membrane is formed by a layer of thick gelatin and sand-piles of calcium carbonate crystals (Fig.l).

Plastic deformation of θ ' in Al-4%Cu alloy

1978 ◽  
Vol 36 (1) ◽  
pp. 576-577
Author(s):  
Shrikant P. Bhat
Keyword(s):  
Deformation Behavior ◽  
Room Temperature ◽  
Electron Microscopic Observation ◽  
Bulk Alloy ◽  
Electron Microscopic ◽  
Cu Alloy ◽  
Cu Alloys ◽  
Orowan Mechanism ◽  
The Subject ◽  
Cyclic Straining

deformation behavior of Al-Cu alloys aged to contain θ ' has been the subject of many investigations (e.g., Ref. 1-5). Since θ ' is strong and hard, dislocations bypass θ ' plates (Orowan mechanism) at low strains. However, at high strains the partially coherent θ ' plates are probably sheared, although the mechanism is complex, depending on the form of deformation. Particularly, the cyclic straining of the bulk alloy is known to produce gross bends and twists of θ '. However, no detailed investigation of the deformation of θ ' has yet been reported; moreover, Calabrese and Laird interpreted the deformation of θ ' as largely being elastic.During an investigation of high temperature cyclic deformation, the detailed electron-microscopic observation revealed that, under reversed straining conditions, θ ' particles are severely distorted--bent and twisted depending on the local matrix constraint. A typical electronmicrograph, showing the twist is shown in Fig. 1. In order to establish whether the deformation is elastic or plastic, a sample from a specimen cycled at room temperature was heated inside the microscope and the results are presented in a series of micrographs (Fig. 2a-e).

Electron Microscopic Observation of Biological Specimens in Their Native State by Employing Cryogenic Techniques

1972 ◽  
Vol 30 ◽  
pp. 410-411 ◽  
Author(s):  
Tokio Nei ◽  
Haruo Yotsumoto ◽  
Yoichi Hasegawa ◽  
Yuji Nagasawa
Keyword(s):  
Electron Microscopic Observation ◽  
Surface Structures ◽  
Specimen Preparation ◽  
Native State ◽  
Electron Microscopic ◽  
Biological Specimen ◽  
Specimen Holder ◽  
Cold Stage ◽  
Preparation Techniques ◽  
Scanning Electron

In order to observe biological specimens in their native state, that is, still containing their water content, various methods of specimen preparation have been used, the principal two of which are the chamber method and the freeze method.Using its recently developed cold stage for installation in the pre-evacuation chamber of a scanning electron microscope, we have succeeded in directly observing a biological specimen in its frozen state without the need for such conventional specimen preparation techniques as drying and metallic vacuum evaporation. (Echlin, too, has reported on the observation of surface structures using the same freeze method.)In the experiment referred to herein, a small sliced specimen was place in the specimen holder. After it was rapidly frozen by freon cooled with liquid nitrogen, it was inserted into the cold stage of the specimen chamber.

Hydrated form of some clay minerals observed using a film sealed environmental cell

1978 ◽  
Vol 36 (1) ◽  
pp. 72-73
Author(s):  
N. Kohyama ◽  
K. Fukushima ◽  
A. Fukami
Keyword(s):  
Clay Minerals ◽  
Morphological Changes ◽  
Carbon Film ◽  
Electron Microscopic Observation ◽  
Adsorbed Water ◽  
Electron Microscopic ◽  
Hydrated Form ◽  
Thin Carbon ◽  
Environmental Cell ◽  
Thin Carbon Film

Since the interlayer or adsorbed water of some clay minerals are quite easily dehydrated in dried air, in vacuum, or at moderate temperatures even in the atmosphere, the hydrated forms have not been observed by a conventional electron microscope(TEM). Recently, specific specimen chambers, “environmental cells(E.C.),” have been developed and confirmed to be effective for electron microscopic observation of wet specimen without dehydration. we observed hydrated forms of some clay minerals and their morphological changes by dehydration using a TEM equipped with an E.C..The E.C., equipped with a single hole copper-microgrid sealed by thin carbon-film, attaches to a TEM(JEM 7A) with an accelerating voltage 100KV and both gas pressure (from 760 Torr to vacuum) and relative humidity can be controlled. The samples collected from various localities in Japan were; tubular halloysite (l0Å) from Gumma Prefecture, sperical halloysite (l0Å) from Tochigi Pref., and intermediate halloysite containing both tubular and spherical types from Fukushima Pref..

Permeability Enhancing and Chemotactic Activities of Lower Molecular Weight Degradation Products of Human Fibrinogen

1981 ◽  
Vol 45 (01) ◽  
pp. 090-094 ◽  
Author(s):  
Katsuo Sueishi ◽  
Shigeru Nanno ◽  
Kenzo Tanaka
Keyword(s):  
Molecular Weight ◽  
Degradation Products ◽  
Electron Microscopic Observation ◽  
Chemotactic Activity ◽  
Lower Molecular Weight ◽  
Electron Microscopic ◽  
Human Fibrinogen ◽  
Rabbit Skin ◽  
Molecular Weights ◽  
Active Fragments

SummaryFibrinogen degradation products were investigated for leukocyte chemotactic activity and for enhancement of vascular permeability. Both activities increased progressively with plasmin digestion of fibrinogen. Active fragments were partially purified from 24 hr-plasmin digests. Molecular weights of the permeability increasing and chemotactic activity fractions were 25,000-15,000 and 25,000 respectively. Both fractions had much higher activities than the fragment X, Y, D or E. Electron microscopic observation of the small blood vessels in rabbit skin correlated increased permeability with the formation of characteristic gaps between adjoining endothelial cells and their contraction.These findings suggest that lower molecular weight degradation products of fibrinogen may be influential in contributing to granulocytic infiltration and enhanced permeability in lesions characterized by deposits of fibrin and/or fibrinogen.

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