scholarly journals Lysine-691 of the anion exchanger from human erythrocytes is located on its cytoplasmic surface

1998 ◽  
Vol 336 (2) ◽  
pp. 443-449 ◽  
Author(s):  
Hans K. ERICKSON ◽  
Jack KYTE
Keyword(s):  
Plasma Membrane ◽  
Anion Exchanger ◽  
Liquid Scintillation ◽  
Pyridoxal Phosphate ◽  
Human Erythrocytes ◽  
Erythrocyte Membranes ◽  
Differential Centrifugation ◽  
Cytoplasmic Surface ◽  
Inside Out

A combination of vectorial modification and site-directed immunochemistry has been used to determine the disposition, with respect to the membrane, of Lys-691 of the anion exchanger from human erythrocytes. Intact erythrocytes and inside-out vesicles were vectorially modified in the same container with pyridoxal phosphate and sodium [3H]borohydride. The modified inside-out vesicles were separated from erythrocytes by differential centrifugation and the vesicles and erythrocyte membranes were treated with alkali and digested with trypsin and thermolysin to liberate the peptides IVSKPER and IVSK{Nε-[4´-(5´-phospho-[4-3H]pyridoxyl)]}PER. These peptides, containing the unmodified and modified versions of Lys-691, were retrieved from the digests by site-directed immunochemistry and were identified by HPLC and liquid scintillation spectroscopy. Both the inside-out vesicles and the intact erythrocytes contained the peptide IVSKPER, however, the 3H-label from the phosphopyridoxylated peptide could be detected only in the inside-out vesicles. The incorporation of 3H into Lys-691 of the anion exchanger from inside-out vesicles was at least 30-fold greater than the incorporation into Lys-691 of the anion exchanger from intact erythrocytes. It follows that Lys-691 of the anion exchanger is located on the cytoplasmic surface of the plasma membrane.

Membrane orientation of sheep spectrin

1980 ◽  
Vol 58 (10) ◽  
pp. 1120-1130
Author(s):  
P. Prokopchuk ◽  
A. U. Sargent
Keyword(s):  
Sheep Erythrocyte ◽  
Human Erythrocytes ◽  
Erythrocyte Membranes ◽  
Variable Degree ◽  
Vertebrate Species ◽  
Membrane Orientation ◽  
Cytoplasmic Surface ◽  
Immunofluorescence Studies ◽  
Surface Labelling ◽  
Inside Out

Based primarily on studies of human erythrocytes, current theories of the structure and organization of erythrocyte membranes localize spectrin to the membrane cytoplasmic surface. Affinity purified anti-sheep spectrin antibodies were used in indirect immunofluorescence studies of intact erythrocytes from various vertebrate species and inside-out and right-side-out impermeable sheep erythrocyte vesicles. This investigation detected i0mmunologically reactive external and potentially transmembranal determinant(s) of the sheep erythrocyte spectrin "assembly." Parallel studies using anti-sheep and anti-human spectrin antibodies, as well as 125I surface-labelling studies of intact sheep and human erythrocytes, indicated that this particular membrane orientation of spectrin was evident in sheep but not in human erythrocytes. Antisera containing antibodies to the external portion of this spectrin "assembly" demonstrated external fluorescence to a variable degree on some, but not all, vertebrate erythrocytes surveyed, confirming that the sheep erythrocyte was not the only exception. It is suggested that there may be subtle species variability in the intermolecular associations of the spectrin "assembly" with(in) the erythrocyte membrane not requiring alterations of the spectrin molecule itself.

scholarly journals Human erythrocytes transport dehydroascorbic acid and sugars using the same transporter complex

2014 ◽  
Vol 306 (10) ◽  
pp. C910-C917 ◽  
Author(s):  
Jay M. Sage ◽  
Anthony Carruthers
Keyword(s):  
Steady State ◽  
Vitamin C ◽  
Blood Cells ◽  
Dehydroascorbic Acid ◽  
Membrane Vesicles ◽  
Human Erythrocytes ◽  
Cytoplasmic Surface ◽  
Glucose Transport Protein ◽  
Inside Out ◽  
Primary Substrate

GLUT1, the primary glucose transport protein in human erythrocytes [red blood cells (RBCs)], also transports oxidized vitamin C [dehydroascorbic acid (DHA)]. A recent study suggests that RBC GLUT1 transports DHA as its primary substrate and that only a subpopulation of GLUT1 transports sugars. This conclusion is based on measurements of cellular glucose and DHA equilibrium spaces, rather than steady-state transport rates. We have characterized RBC transport of DHA and 3- O-methylglucose (3-OMG), a transported, nonmetabolizable sugar. Steady-state 3-OMG and DHA uptake in the absence of intracellular substrate are characterized by similar Vmax (0.16 ± 0.01 and 0.13 ± 0.02 mmol·l−1·min−1, respectively) and apparent Km (1.4 ± 0.2 and 1.6 ± 0.7 mM, respectively). 3-OMG and DHA compete for uptake, with Ki(app) of 0.7 ± 0.4 and 1.1 ± 0.1 mM, respectively. Uptake measurements using RBC inside-out-membrane vesicles demonstrate that 3-OMG and DHA compete at the cytoplasmic surface of the membrane, with Ki(app) of 0.7 ± 0.1 and 0.6 ± 0.1 mM, respectively. Intracellular 3-OMG stimulates unidirectional uptake of 3-OMG and DHA. These findings indicate that DHA and 3-OMG bind at mutually exclusive sites at exo- and endofacial surfaces of GLUT1 and are transported via the same GLUT1 complex.

scholarly journals Spectrin-dependent and -independent association of F-actin with the erythrocyte membrane.

1980 ◽  
Vol 86 (2) ◽  
pp. 694-698 ◽  
Author(s):  
C M Cohen ◽  
S F Foley
Keyword(s):  
Membrane Protein ◽  
Erythrocyte Membrane ◽  
Actin Filaments ◽  
Binding Capacity ◽  
Protein Band ◽  
Actin Binding ◽  
Erythrocyte Membranes ◽  
Cytoplasmic Surface ◽  
Independent Association ◽  
Inside Out

Binding of F-actin to spectrin-actin-depleted erythrocyte membrane inside-out vesicles was measured using [3H]F-actin. F-actin binding to vesicles at 25 degrees C was stimulated 5-10 fold by addition of spectrin dimers or tetramers to vesicles. Spectrin tetramer was twice as effective as dimer in stimulating actin binding, but neither tetramer nor dimer stimulated binding at 4 degrees C. The addition of purified erythrocyte membrane protein band 4.1 to spectrin-reconstituted vesicles doubled their actin-binding capacity. Trypsinization of unreconstituted vesicles that contain < 10% of the spectrin but nearly all of the band 4.1, relative to ghosts, decreased their F-actin-binding capacity by 70%. Whereas little or none of the residual spectrin was affected by trypsinization, band 4.1 was significantly degraded. Our results show that spectrin can anchor actin filaments to the cytoplasmic surface of erythrocyte membranes and suggest that band 4.1 may be importantly involved in the association.

Carboxyl-Terminal Truncations of Human Anion Exchanger Impair its Trafficking to the Plasma Membrane

Traffic ◽  
2003 ◽  
Vol 4 (9) ◽  
pp. 642-651 ◽  
Author(s):  
Emmanuelle Cordat ◽  
Jing Li ◽  
Reinhart A. F. Reithmeier
Keyword(s):  
Plasma Membrane ◽  
Anion Exchanger ◽  
Carboxyl Terminal

scholarly journals Influence of the glycocalyx and plasma membrane composition on amphiphilic gold nanoparticle association with erythrocytes

Nanoscale ◽  
2015 ◽  
Vol 7 (26) ◽  
pp. 11420-11432 ◽  
Author(s):  
Prabhani U. Atukorale ◽  
Yu-Sang Yang ◽  
Ahmet Bekdemir ◽  
Randy P. Carney ◽  
Paulo J. Silva ◽  
...  
Keyword(s):  
Gold Nanoparticles ◽  
Plasma Membrane ◽  
Gold Nanoparticle ◽  
Erythrocyte Membranes ◽  
Membrane Composition ◽  
Membrane Components

Amphiphilic gold nanoparticles spontaneously insert into erythrocyte membranes; we characterize this association as a function of key plasma membrane components.

scholarly journals A carbohydrate-deficient membrane glycoprotein in human erythrocytes of phenotype S-s-

1977 ◽  
Vol 165 (1) ◽  
pp. 157-161 ◽  
Author(s):  
M J A Tanner ◽  
D J Anstee ◽  
P A Judson
Keyword(s):  
Blood Group ◽  
Arachis Hypogaea ◽  
Human Erythrocyte ◽  
Structure And Function ◽  
Human Erythrocytes ◽  
Erythrocyte Membranes ◽  
Membrane Glycoprotein ◽  
Blood Group Antigens ◽  
And Function ◽  
Normal Erythrocyte

1. We investigated the membranes of human erythrocytes which completely lack the blood-group antigens S and s (denoted as S-s-) as part of a study of the structure and function of the surface glycoproteins of the human erythrocyte. 2. The S-s-erythrocyte-membrane glycoprotein PAS-3 band was much less intensely stained in comparison with that of the glycoprotein from normal erythrocyte membranes. The S-s-membrane glycoprotein PAS-4 band also showed decreased staining. 3. Examination with the lectins from Maclura aurantiaca (Osage orange) and Arachis hypogaea (groundnut) showed that the PAS-3 glycoprotein of S-s-erythrocyte membranes lacked the receptors for these lectins that are present on glycoprotein PAS-3 from normal erythrocytes. 4. Radioiodination with lactoperoxidase showed the presence of the polypeptide of glycoprotein PAS-3 in S-s-cells, although it was more weakly labelled than the protein in the normal erythrocyte. 5. Our results show that the PAS-3 glycoprotein of S-s-erythrocytes is deficient in some of the carbohydrates present in the protein from normal erythrocytes. Glycoprotein PAS-4 of normal erythrocytes is shown to be a complex containing both glycoproteins PAS-1 and PAS-3.

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