Polyribosomes from L cells

Dowben, Robert M.; Lynch, P.M.; Nadler, Henry L.; Hsia, David Yi‐Yung

doi:10.1016/0014-4827(69)90128-1

Cited by 16 publications

(4 citation statements)

References 7 publications

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“…Rough microsomes are particularly susceptible to shearing forces (29) resulting in the release of ribosomes from membranes, and it was thus considered essential to eliminate this possibility in studies on the role of the endoplasmic reticulum in protein synthesis. Nitrogen cavitation (30) has been used by Dowben et aL (6,7) for isolating polysomes from various cultured cells and animal tissue. They concluded that nitrogen cavitation results in complete disrup-tion of almost all cells with minimal damage to the polyribosomes.…”

Section: Discussionmentioning

confidence: 99%

“…In order to examine more closely the membranebound polysome population it was necessary to introduce a technique that allowed for disruption of cells without the use of detergents, since an initial solubilization of endoplasmic reticulum membranes had to be avoided. The technique of nitrogen cavitation has been found to be extremely useful in the isolation of large amounts of intact 'heavy' polysomes from various types of cells (6,7,8), and this method of causing cell disruption was thus adopted. Initially this enabled the separation of two populations of membrane-bound polysomesthose of the rough microsomes and those of the nuclear-associated endoplasmic reticulum (9).…”

Section: Introductionmentioning

confidence: 99%

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Ultrastructure and polysome content of the microsomal subfractions of mouse plasmacytoma cells

1981

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The endoplasmic reticulum (ER) of MPC-11 cells released as vesicles upon cell disruption by nitrogen cavitation was separated from the bulk of mitochondria, lysosomes and plasma membranes by a low speed centrifugation. The ER membranes were fractionated on discontinuous sucrose gradients into heavy rough (HR), light rough (LR) and smooth (S) membranes. The morphology of subcellular fractions was studied by electron microscopy and the ER membranes were shown to be virtually free of contaminating organelles. The S fraction was easily distinguishable because of the lack or ribosomes but there were no apparent morphological differences between the HR and LR fractions. Of total activity in the microsomal subfractions, 70% of the UDPase and 67% of the 5'-nucleotidase activity was associated with the S fraction. Polysomes were present in the HR, LR and nuclear-associated ER fractions but not in the S fraction. The HR and LR fractions did not appear to be contaminated to any great extent with free polysomes. RNA/protein and RNA/phospholipid ratios of the HR fraction were higher than those of the LR fraction, indicating a greater density of ribosomes in the former fraction. These ratios were much lower in the S fraction reflecting the low ribosome content.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Ultrastructure and polysome content of the microsomal subfractions of mouse plasmacytoma cells

1981

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“…For oligomeric protein structures, these subunits are rapidly assembled into receptors and queued for transport to the plasma membrane (PM) 10. In order to isolate single receptors expressed in these cell membranes, we fragmented cells by using nitrogen cavitation with a pressure of approximately 250 psi for 5 min to form membrane vesicles (see the Supporting Information) 11. The resulting individual membrane fragments spontaneously reform to create vesicles.…”

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confidence: 99%

Evaluation of the Efficiency of Filtration Processes Using Precoat Materials

2016

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The highly efficient filtration of ultrafine dust emitted by biomass combustion processes with a baghouse filter has been successfully tested in the last years. To protect the filter material from the very small and sticky fine dust particles and to guarantee very high total collection efficiencies (> 99 %) in a long‐term stable process, the use of a precoat is necessary. Tests done in a laboratory and a real‐application plant show that the reuse of precoat materials can lead to significant savings. Considering the influences of different combustion processes, the associated precoat efficiencies could be calculated. With these characteristic ratios, it is possible to evaluate different process settings. Hence, the amount and the cost of the needed precoat could be reduced significantly.

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“…[10] In order to isolate single receptors expressed in these cell membranes, we fragmented cells by using nitrogen cavitation with a pressure of approximately 250 psi for 5 min to form membrane vesicles (see the Supporting Information). [11] The resulting individual membrane fragments spontaneously reform to create vesicles. Vesicles were separated from the cell lysate through differential ultracentrifugation.…”

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confidence: 99%

Cell‐Derived Vesicles for Single‐Molecule Imaging of Membrane Proteins

et al. 2014

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A new approach is presented for the application of single-molecule imaging to membrane receptors through the use of vesicles derived from cells expressing fluorescently labeled receptors. During the isolation of vesicles, receptors remain embedded in the membrane of the resultant vesicles, thus allowing these vesicles to serve as nanocontainers for single-molecule measurements. Cell-derived vesicles maintain the structural integrity of transmembrane receptors by keeping them in their physiological membrane. It was demonstrated that receptors isolated in these vesicles can be studied with solution-based fluorescence correlation spectroscopy (FCS) and can be isolated on a solid substrate for single-molecule studies. This technique was applied to determine the stoichiometry of α3β4 nicotinic receptors. The method provides the capability to extend single-molecule studies to previously inaccessible classes of receptors.

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Polyribosomes from L cells

Cited by 16 publications

References 7 publications

Ultrastructure and polysome content of the microsomal subfractions of mouse plasmacytoma cells

Ultrastructure and polysome content of the microsomal subfractions of mouse plasmacytoma cells

Evaluation of the Efficiency of Filtration Processes Using Precoat Materials

Cell‐Derived Vesicles for Single‐Molecule Imaging of Membrane Proteins

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