Variations in distribution of con A receptor sites and anionic groups during red blood cell differentiation in the rat.

Skutelsky, Ehud; Farquhar, Marilyn G.

doi:10.1083/jcb.71.1.218

Cited by 85 publications

(40 citation statements)

References 36 publications

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“…For example, during the enucleation of rat erythroblasts in vivo, receptors for the lectin concanavalin A are preferentially lost from the cell with the extruded nucleus, whereas the anionic receptors for colloidal iron hydroxide are preferentially retained in the reticulocyte portion of the cell. The major colloidal iron hydroxide-binding glycoproteins of the mature erythrocyte are the glycophorin molecules (26), which bear most of the surface sialic acid. These results, therefore, indicate the selective retention of glycophorin in the reticulocyte during enucleation.…”

Section: Resultsmentioning

confidence: 99%

Membrane protein redistribution during differentiation of cultured human erythroleukemic cells.

Hunt¹,

Marshall²

1981

Mol. Cell. Biol.

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Human erythroleukemic (K562) cells differentiate along the erythroid differentiation pathway in vitro when 0.05 mM hemin is included in the growth medium. In the presence of the inducer the cells continue to proliferate and, after a delay of 24 to 48 h, start to synthesize hemoglobin. However, during differentiation, no changes in the major cell surface proteins were detected using lactoperoxidase-catalyzed iodination, and no change in the synthesis of spectrin, the major cytoskeletal protein of the mature erythrocyte, was detected by specific immune precipitation. Despite this absence of major changes in cell surface proteins, profound changes take place in the organization of the cell membrane. A process similar but not identical to the enucleation observed in erythroid differentiation in vivo occurs in which a smooth-surfaced cell, about 10 jim in diameter, is divided from the nucleus-containing part of the cell. With the exception of ribosomes, these reticulocyte-like cells contain no organelles when examined by transmission electron microscopy, but contain much of the parent cell's hemoglobin, spectrin, and glycophorin.The surface membrane (or ghost) of the human erythrocyte is undoubtedly the best-characterized eucaryotic cell membrane, and a considerable amount of information concerning the structure of its major proteins has accumulated (for a review see reference 23). The major integral membrane glycoproteins of the erythrocyte membrane, glycophorin (which bears the MN antigens [27]) and band 3 protein (which is the major anion transport channel [6]), are located on the external surface of the cell, span the lipid bilayer (3, 28), and interact with peripheral proteins on the cytoplasmic side of the membrane (for a review see reference 23). Of these peripheral proteins the most actively studied have been spectrin (bands 1 and 2 in the nomenclature of Fairbanks and colleagues [10]) and actin (band 5), which together comprise the cytoskeleton lining the inner surface of the membrane (20). It is this cytoskeleton that is responsible for the cells' biconcave shape and spectacular deformability (21 Transmission electron microscopy. Cells were fixed using 2% glutaraldehyde in 0.1 M sodium cacodylate buffer (pH 7.4). They were then incubated with 1% osmium tetroxide in water and processed for electron microscopy using uranyl acetate and lead citrate staining.Scanning electron microscopy. Cells were washed in cold PBS and suspended in 2% glutaraldehyde in PBS at 4°C. After 24 h the cells were gently resuspended, and 1 drop of the suspension was placed on the shiny side of a Nucleopore polycarbonate filter (0.4-um pore size; 13-mm diameter) supported on the grid half of a Gelman Easy-pressure filter holder attached to a 5-ml syringe. The cells were gently drawn down onto the filter and fixed on the filter with 2% glutaraldehyde in PBS. The cells and filter were postfixed in PBS containing 1% osmium tetroxide for 1 h at room temperature and then dehydrated in ethanol, followed by critical-point drying. The filters...

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Section: Resultsmentioning

confidence: 99%

Membrane protein redistribution during differentiation of cultured human erythroleukemic cells.

Hunt¹,

Marshall²

1981

Mol. Cell. Biol.

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“…Other instances of membrane polarization in cells devoid of tight junctions exemplify that lateral diffusion of proteins may be prevented in the absence of junctional complexes. Among these are the erythroblast at the time of nuclear expulsion (38) and adherent macrophages (47), a cell type possibly related with the osteoclast. We propose that the sealing zone, where actin filaments are anchored to the membrane and the cell is attached to its substratum (25), could play such a restrictive role.…”

Section: Discussionmentioning

confidence: 99%

Cell-mediated extracellular acidification and bone resorption: evidence for a low pH in resorbing lacunae and localization of a 100-kD lysosomal membrane protein at the osteoclast ruffled border.

Baron¹,

Neff²,

Louvard³

et al. 1985

The Journal of Cell Biology

690

332

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The extracellular compartment where bone resorption occurs, between the osteoclast and bone matrix, is shown in this report to be actively acidified. The weak base acridine orange accumulates within this compartment but dissipates after incubation with ammonium chloride. Upon removal of ammonium chloride, the cells are able to rapidly reacidify this compartment. The highly convoluted plasma membrane of the osteoclast facing this acidic compartment (ruffled border) is shown to contain a 100-kD integral membrane protein otherwise present in limiting membranes of lysosomes and other related acidified organelles

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“…3,[8][9][10] Earlier studies also report that nonsialated glycoproteins are enriched in membranes of extruded nuclei, while sialoglycoproteins are enriched in membranes of young reticulocytes. 11 However, the redistribution of a large number of well-characterized integral membrane proteins and the mechanism(s) underlying their redistribution are unexplored.…”

Section: Introductionmentioning

confidence: 99%

Mechanism of protein sorting during erythroblast enucleation: role of cytoskeletal connectivity

Lee

Gimm

et al. 2004

Blood

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During erythroblast enucleation, nuclei surrounded by plasma membrane separate from erythroblast cytoplasm. A key aspect of this process is sorting of erythroblast plasma membrane components to reticulocytes and expelled nuclei. Although it is known that cytoskeletal elements actin and spectrin partition to reticulocytes, little is understood about molecular mechanisms governing plasma membrane protein sorting. We chose glycophorin A (GPA) as a model integral protein to begin investigating proteinsorting mechanisms. Using immunofluorescence microscopy and Western blotting we found that GPA sorted predominantly to reticulocytes. We hypothesized that the degree of skeletal linkage might control the sorting pattern of transmembrane proteins. To explore this hypothesis, we quantified the extent of GPA association to the cytoskeleton in erythroblasts, young reticulocytes, and mature erythrocytes using fluorescence imaged microdeformation (FIMD) and observed that GPA underwent dramatic reorganization during terminal differentiation. We discovered that GPA was more connected to the membrane cytoskeleton, either directly or indirectly, in erythroblasts and young reticulocytes than in mature cells. We conclude that skeletal protein association can regulate protein sorting during enucleation. Further, we suggest that the enhanced rigidity of reticulocyte membranes observed in earlier investigations results, at least in part, from increased connectivity of GPA with the spectrinbased skeleton. IntroductionDuring mammalian erythroid terminal differentiation, the plasma membrane and cytoskeleton are in a state of dynamic reorganization. We and others have determined that changes in protein expression and membrane protein assembly occur that involve both integral and skeletal membrane components. [1][2][3][4][5][6][7] At the conclusion of terminal differentiation, erythroblasts expel their nuclei and become reticulocytes. During enucleation, nuclei surrounded by plasma membrane separate from erythroblast cytoplasm. A key aspect of this process is the sorting of erythroblast plasma membrane components to the plasma membranes of the nascent reticulocyte and the expelled nucleus. Although approximately 2 million reticulocytes are generated each second, amazingly little is known about molecular mechanisms governing protein sorting during enucleation. It is known that actin, spectrin, tubulin, ankyrin, and protein 4.1 partition to young reticulocytes, leaving extruded nuclei devoid of skeletal elements. 3,[8][9][10] Earlier studies also report that nonsialated glycoproteins are enriched in membranes of extruded nuclei, while sialoglycoproteins are enriched in membranes of young reticulocytes. 11 However, the redistribution of a large number of well-characterized integral membrane proteins and the mechanism(s) underlying their redistribution are unexplored.Since glycophorin A (GPA) is a well-defined, major sialoglycoprotein in the mature erythrocyte (as reviewed in Chasis and Mohandas 12 ), we chose it as a model integral membrane ...

show abstract

Variations in distribution of con A receptor sites and anionic groups during red blood cell differentiation in the rat.

Cited by 85 publications

References 36 publications

Membrane protein redistribution during differentiation of cultured human erythroleukemic cells.

Membrane protein redistribution during differentiation of cultured human erythroleukemic cells.

Cell-mediated extracellular acidification and bone resorption: evidence for a low pH in resorbing lacunae and localization of a 100-kD lysosomal membrane protein at the osteoclast ruffled border.

Mechanism of protein sorting during erythroblast enucleation: role of cytoskeletal connectivity

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