Role of plasma membrane and of cytomatrix in maintenance of intracellular to extracellular ion gradients in chicken erythrocytes

Cameron, Ivan L.; Hunter, Keithley E.; Smith, Nancy R.; Hazlewood, Carlton F.; Ludány, Andrea; Kellermayer, Miklós

doi:10.1002/jcp.1041370213

Cited by 11 publications

(4 citation statements)

References 13 publications

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“…Other experiments, employing high-voltage electron microscopy and cell extraction procedures, demonstrated the presence of an organized network in cells (22,23) which might act as a scaffold for cell organization (20). Subsequent work revealed that some glycolytic enzymes (5) and some detergent-extractable proteins (2) are not freely diffusible in vivo, suggesting that at least some cellular components might be present in highly organized structures (reviewed in reference 26).…”

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confidence: 99%

Organization of Mammalian Cytoplasm

Hudder

Nathanson

Deutscher

2003

Molecular and Cellular Biology

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Although the role of macromolecular interactions in cell function has attracted considerable attention, important questions about the organization of cells remain. To help clarify this situation, we used a simple protocol that measures macromolecule release after gentle permeabilization for the examination of the status of endogenous macromolecules. Treatment of Chinese hamster ovary cells with saponin under carefully controlled conditions allowed entry of molecules of at least 800 kDa; however, there were minimal effects on internal cellular architecture and protein synthesis remained at levels comparable to those seen with intact cells. Most importantly, total cellular protein and RNA were released from these cells extremely slowly. The release of actin-binding proteins and a variety of individual cytoplasmic proteins mirrored that of total protein, while marker proteins from subcellular compartments were not released. In contrast, glycolytic enzymes leaked rapidly, indicating that cells contain at least two distinct populations of cytoplasmic proteins. Addition of microfilament-disrupting agents led to rapid and extensive release of cytoplasmic macromolecules and a dramatic reduction in protein synthesis. These observations support the conclusion that mammalian cells behave as highly organized, macromolecular assemblies (dependent on the actin cytoskeleton) in which endogenous macromolecules normally are not free to diffuse over large distances.Tremendous progress has been made in our understanding of cell function. For the most part, this has been accomplished through the use of a traditional reductionist approach in which individual cellular components are identified and isolated and their cellular roles are reconstructed on the basis of their functions in vitro. While such an approach has proven to be highly successful, especially for determining the "players" in cell metabolism, it falls short in explaining how these components actually function within the cell. In fact, in many cases, particularly those involving complex cellular processes, it often has not been possible to recreate the efficiency of cellular reactions in vitro. Understanding what accounts for such differences in efficiency is essential if we are to explain cellular function in its entirety.In recent years, considerable attention has focused on the importance of macromolecular interactions in cell function (see, e.g., reference 10) and on the fact that enzymes contributing to complex processes often are bound to each other and that intermediates in the process may be channeled (see, e.g., references 6 and 16 and the review in reference 19). As a consequence of such organization, processes within cells may be able to proceed much more efficiently than those carried out by the same enzymes dispersed in solution in vitro. Thus, important questions that remain to be answered are (i) how extensive is cellular organization, (ii) what cellular components are responsible for maintaining it, and (iii) are macromolecular interactions confined to i...

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confidence: 99%

Organization of Mammalian Cytoplasm

Hudder

Nathanson

Deutscher

2003

Molecular and Cellular Biology

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“…The sudden disorganization of this cooperative association would result in mobilization of Hb. In this regard Brij-permeabilized chicken erythrocytes also showed evidence of a sudden loss of Hb from individual erythrocytes (Cameron et al, 1988a). Because a Brij-resistent pervasive cytoskeleton (not a continuous submembrane lamina) remained in the chicken erythrocyte, the sudden rupture of a submembrane lamina can be ruled out as an explanation for the sudden loss of Hb from the chicken erythrocyte.…”

Section: Assessment Of the Mobility Of Hemoglobin And Heavy-water Holmentioning

confidence: 96%

Maintenance and mobility of hemoglobin and water within the human erythrocyte after detergent disruption of the plasma membrane

Cameron

Cox

Liu

et al. 1991

Journal Cellular Physiology

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Is an intact plasma membrane responsible for keeping hemoglobin and water within the human erythrocyte? If not, what is responsible? How free is Hb to move about within the erythrocyte? To answer these questions erythrocytes were taken for phase contrast microscopy, transmission electron microscopy (TEM), determination of water-holding capacity, and proton NMR studies both before and after membrane disruption with a nonionic detergent (Brij 58). Addition of 0.2% Brij to a D2O saline solution of hemoglobin (Hb) caused particles of Hb to appear and to aggregate. This aggregation of Hb caused the amplitude of the Hb proton NMR spectra to decrease. Thus, the less mobile the Hb the lower the Hb proton spectra amplitude. Erythrocytes washed in D2O saline showed proton NMR spectra of relatively low amplitude. Addition of Brij (0.2%) to these erythrocytes caused increased Hb mobility within these erythrocytes. The TEM of fixed and thin-sectioned erythrocytes treated with Brij showed disruption of the plasma membrane of all erythrocytes regardless of whether or not they had lost Hb. Brij-permeabilized erythrocytes washed in D2O saline or in a D2O K buffer maintained a higher heavy water-holding capacity upon centrifugation as compared to nonpermeabilized erythrocytes. The TEM of Brij-treated and washed erythrocyte "shells" revealed a continuous submembrane lamina but no other evidence of cytoskeletal elements. The water-holding capacity of the erythrocyte can be accounted for by the water-holding capacity of hemoglobin. The evidence favors a relatively immobile state of Hb and of water in the erythrocyte that is not immediately dependent on an intact plasma membrane but is attributed to interactions between Hb molecules and the submembrane lamina.

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“…Plasma membrane, endosomes (early or late), and lysosomes are dynamic organelles that are known for their key role in cellular function such as cell signaling [ 1 – 3 ], intracellular [ 4 , 5 ] and extracellular interaction [ 6 , 7 ]. Interestingly, most drugs and therapeutic small molecules require cellular interaction for their biological action, and it is well known that high affinity with the cell surface results in effective mechanisms [ 8 ].…”

Section: Introductionmentioning

confidence: 99%

Surface functionalization dependent subcellular localization of Superparamagnetic nanoparticle in plasma membrane and endosome

Raj

Khan²

2018

Nano Convergence

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In this article, we elaborate the application of thermal decomposition based synthesis of Fe3O4 superparamagnetic nanoparticle (SPMNP) in subcellular fractionation context. Here, we performed surface functionalization of SPMNP with phospholipids and dimercaptosuccinic acid. Surprisingly, we observed surface functionalization dependent SPMNP localization in subcellular compartments such as plasma membrane, endosomes and lysosomes. By using SPMNP based subcellular localization with pulse–chase methodology, we could use SPMNP for high pure-high yield organelle (plasma membrane, endosomes and lysosome) fractionation. Further, SPMNP that are distinctly localized in subcellular compartments can be used as technology for subcellular fractionation that can complement existing tools for cell biology research. As a future perspective, isolated magnetic organelles can be extended to protein/protein complex purification for biochemical and structural biology studies.

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Role of plasma membrane and of cytomatrix in maintenance of intracellular to extracellular ion gradients in chicken erythrocytes

Cited by 11 publications

References 13 publications

Organization of Mammalian Cytoplasm

Organization of Mammalian Cytoplasm

Maintenance and mobility of hemoglobin and water within the human erythrocyte after detergent disruption of the plasma membrane

Surface functionalization dependent subcellular localization of Superparamagnetic nanoparticle in plasma membrane and endosome

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