scholarly journals Metabolic dependence of protein arrangement in human erythrocyte membranes. I. Analysis of spectrin-rich complexes in ATP-depleted red cells

Blood ◽  
1978 ◽  
Vol 51 (3) ◽  
pp. 385-395 ◽  
Author(s):  
J Palek ◽  
SC Liu ◽  
LM Snyder
Keyword(s):  
Human Erythrocyte ◽  
Polyacrylamide Gel Electrophoresis ◽  
Band 3 ◽  
Atp Depletion ◽  
Red Cells ◽  
Erythrocyte Membranes ◽  
Red Cell ◽  
Aerobic Conditions ◽  
Spontaneous Formation ◽  
Human Erythrocyte Membranes

Abstract The discocyte-echinocyte transformation and the decrease in deformability associated with red cell ATP depletion have been attributed to changes in the physical properties of spectrin and actin, membrane proteins located at the membrane-cytosol interface. We investigated the spontaneous formation of spectrin-rich complexes in human erythrocyte membranes, employing two-dimensional SDS- polyacrylamide gel electrophoresis. Membranes of red cells depleted in ATP under aerobic conditions exhibited (1) an increase in components 4.5 and 8 and globin subunits, (2) a spontaneous formation of heterodimers of spectrin 1 + 2 and spectrin 2 + component 4.9, and (3) a large molecular weight (greater than 10(6) daltons) protein complex with a high spectrin to band 3 ratio. These complexes were dissociated with dithiothreitol and were prevented by anaerobic incubation or the maintenance of red cell ATP and GSH levels with glucose, adenine, and inosine. The complexes 1 + 2 and 2 + 4.9 were also seen in acetylphenylhydrazine-treated, glucose-6-phosphate dehydrogenase- deficient fresh erythrocytes that showed marked GSH depletion but preserved greater than 70% of the original ATP level. However, membranes of these cells did not contain the greater 10(6) dalton aggregate with a high spectrin to band 3 ratio. We concluded that the formation of the latter complex results from rearrangement of spectrin and other polypeptides in membranes of ATP-depleted red cells. Under aerobic conditions, the rearranged proteins undergo spontaneous intermolecular crosslinkings through disulfide couplings.

scholarly journals Metabolic dependence of protein arrangement in human erythrocyte membranes. I. Analysis of spectrin-rich complexes in ATP-depleted red cells

Blood ◽  
1978 ◽  
Vol 51 (3) ◽  
pp. 385-395
Author(s):  
J Palek ◽  
SC Liu ◽  
LM Snyder
Keyword(s):  
Human Erythrocyte ◽  
Polyacrylamide Gel Electrophoresis ◽  
Band 3 ◽  
Atp Depletion ◽  
Red Cells ◽  
Erythrocyte Membranes ◽  
Red Cell ◽  
Aerobic Conditions ◽  
Spontaneous Formation ◽  
Human Erythrocyte Membranes

The discocyte-echinocyte transformation and the decrease in deformability associated with red cell ATP depletion have been attributed to changes in the physical properties of spectrin and actin, membrane proteins located at the membrane-cytosol interface. We investigated the spontaneous formation of spectrin-rich complexes in human erythrocyte membranes, employing two-dimensional SDS- polyacrylamide gel electrophoresis. Membranes of red cells depleted in ATP under aerobic conditions exhibited (1) an increase in components 4.5 and 8 and globin subunits, (2) a spontaneous formation of heterodimers of spectrin 1 + 2 and spectrin 2 + component 4.9, and (3) a large molecular weight (greater than 10(6) daltons) protein complex with a high spectrin to band 3 ratio. These complexes were dissociated with dithiothreitol and were prevented by anaerobic incubation or the maintenance of red cell ATP and GSH levels with glucose, adenine, and inosine. The complexes 1 + 2 and 2 + 4.9 were also seen in acetylphenylhydrazine-treated, glucose-6-phosphate dehydrogenase- deficient fresh erythrocytes that showed marked GSH depletion but preserved greater than 70% of the original ATP level. However, membranes of these cells did not contain the greater 10(6) dalton aggregate with a high spectrin to band 3 ratio. We concluded that the formation of the latter complex results from rearrangement of spectrin and other polypeptides in membranes of ATP-depleted red cells. Under aerobic conditions, the rearranged proteins undergo spontaneous intermolecular crosslinkings through disulfide couplings.

The nature of human erythrocyte receptors for Neisseria gonorrhoeae

1982 ◽  
Vol 28 (2) ◽  
pp. 219-222 ◽  
Author(s):  
G. M. Wiseman ◽  
P. McNicol
Keyword(s):  
Neisseria Gonorrhoeae ◽  
Human Erythrocyte ◽  
Polyacrylamide Gel Electrophoresis ◽  
Protein Band ◽  
Band 3 ◽  
Erythrocyte Membranes ◽  
Glycophorin A ◽  
Receptor Sites ◽  
Human Erythrocyte Membranes ◽  
Sepharose 4B

Normal and trypsinized human erythrocyte membranes were used as a model in the study of host cell receptors for Neisseria gonorrhoeae. Receptor sites were identified by adherence inhibition assays of fractions of membranes eluted from polyacrylamide gel electrophoresis columns. Results indicated that inhibition of gonococcus T1 and T4 adherence was associated with erythrocyte protein bands 3 and 4 and glycophorin A, the major sialoglycoprotein. Further investigation revealed that band 3 preparations isolated by affinity chromatography on concanavalin A – Sepharose 4B columns continued to inhibit T1 adherence to erythrocytes but did not inhibit adherence of T4 organisms. It is suggested that protein band 3 is the receptor on erythrocytes for T1 gonococci and that glycophorin A may be the receptor for T4 cells.

scholarly journals Ultrastructure of the intact skeleton of the human erythrocyte membrane.

1986 ◽  
Vol 102 (3) ◽  
pp. 997-1006 ◽  
Author(s):  
B W Shen ◽  
R Josephs ◽  
T L Steck
Keyword(s):  
Human Erythrocyte ◽  
Actin Filaments ◽  
Band 3 ◽  
Carbon Films ◽  
Erythrocyte Membranes ◽  
Accessory Proteins ◽  
Red Cell Membrane ◽  
Human Erythrocyte Membranes ◽  
Low Ionic Strength ◽  
Triton X

Filamentous skeletons were liberated from isolated human erythrocyte membranes in Triton X-100, spread on fenestrated carbon films, negatively stained, and viewed intact and unfixed in the transmission electron microscope. Two forms of the skeleton were examined: (a) basic skeletons, stripped of accessory proteins with 1.5 M NaCl so that they contain predominantly polypeptide bands 1, 2, 4.1, and 5; and (b) unstripped skeletons, which also bore accessory proteins such as ankyrin and band 3 and small plaques of residual lipid. Freshly prepared skeletons were highly condensed. Incubation at low ionic strength and in the presence of dithiothreitol for an hour or more caused an expansion of the skeletons, which greatly increased the visibility of their elements. The expansion may reflect the opening of spectrin from a compact to an elongated disposition. Expanded skeletons appeared to be organized as networks of short actin filaments joined by multiple (5-8) spectrin tetramers. In unstripped preparations, globular masses were observed near the centers of the spectrin filaments, probably corresponding to complexes of ankyrin with band 3 oligomers. Some of these globules linked pairs of spectrin filaments. Skeletons prepared with a minimum of perturbation had thickened actin protofilaments, presumably reflecting the presence of accessory proteins. The length of these actin filaments was highly uniform, averaging 33 +/- 5 nm. This is the length of nonmuscle tropomyosin. Since there is almost enough tropomyosin present to saturate the F-actin, our data support the hypothesis that tropomyosin may determine the length of actin protofilaments in the red cell membrane.

Isolation and Partial Structure of an Oligosaccharide of Band-3 Glycoproteins of Human Erythrocyte Membranes

1979 ◽  
pp. 1065-1072
Author(s):  
Toshiaki Osawa ◽  
Tsutomu Tsuji ◽  
Anne Marianne Golovtchenko-Matsumoto ◽  
Tatsuro Irimura
Keyword(s):  
Human Erythrocyte ◽  
Band 3 ◽  
Erythrocyte Membranes ◽  
Partial Structure ◽  
Human Erythrocyte Membranes

Protoporphyrin-induced photodynamic effects on band 3 protein of human erythrocyte membranes

1981 ◽  
Vol 649 (2) ◽  
pp. 310-316 ◽  
Author(s):  
T.M.A.R. Dubbelman ◽  
A.F.P.M. De Goeij ◽  
K. Christianse ◽  
J. Van Steveninck
Keyword(s):  
Human Erythrocyte ◽  
Band 3 ◽  
Erythrocyte Membranes ◽  
Band 3 Protein ◽  
Human Erythrocyte Membranes

scholarly journals Degradation of Band-3 Glycoprotein in vitro by a Protease Isolated from Human Erythrocyte Membranes

1982 ◽  
Vol 121 (2) ◽  
pp. 463-467 ◽  
Author(s):  
Anne Marianne GOLOVTCHENKO-MATSUMOTO ◽  
Isamu MATSUMOTO ◽  
Toshiaki OSAWA
Keyword(s):  
Human Erythrocyte ◽  
Band 3 ◽  
Erythrocyte Membranes ◽  
Human Erythrocyte Membranes

scholarly journals ATP11C Encodes a Major Flippase in Human Erythrocyte and Its Genetic Defect Causes Congenital Non-Spherocytic Hemolytic Anemia

Blood ◽  
2015 ◽  
Vol 126 (23) ◽  
pp. 2131-2131
Author(s):  
Nobuto Arashiki ◽  
Yuichi Takakuwa ◽  
Hiromi Ogura ◽  
Taiju Utsugisawa ◽  
Satoru Miyano ◽  
...  
Keyword(s):  
Hemolytic Anemia ◽  
Human Erythrocyte ◽  
Conflicts Of Interest ◽  
Massively Parallel Sequencing ◽  
Genetic Defect ◽  
Red Cells ◽  
Single Amino Acid ◽  
Erythrocyte Membranes ◽  
Splenic Macrophages ◽  
Human Erythrocyte Membranes

Abstract Flippases are members of the P-IV ATPase family of proteins, and contribute to localization of phosphatidylserine (PS) in inner leaflet by its ATP-dependent active transport from outer to inner leaflet of lipid bilayer of erythrocyte membranes. It is critical that erythrocytes maintain PS in the inner monolayer to ensure their 120-day survival in circulation since externalization of PS will be recognized as an eat-me signal, resulting in phagocytosis by splenic macrophages. In the present study, we identified that ATP11C gene encodes a major flippase in human erythrocytes. A 13 years-old boy was referred to our hospital for consultation for work up of an undiagnosed congenital hemolytic anemia. Since extensive biochemical and molecular analysis failed to identify hemoglobin, erythrocyte membranes and enzyme abnormalities for the pathogenesis of hemolysis, we performed the whole exome analysis by massively parallel sequencing. We identified that the proband is hemizyogous, and the mother is heterozygous for a missense mutation of ATP11C, c.1253C>A, corresponding to a single amino acid substitution, p.Thr418Asn. Flipping activity as measured by PS internalization was decreased to 10% in the red cells of the proband compared to a normal control, clearly demonstrating that ATP11C encodes a major flippase in the human erythrocyte membranes. The PS-positive erythrocytes were not significantly increased in the whole blood but only in the most dense senescent cells, suggesting that PS exposure did not occur until very late stages of lifespan. We showed that PS exposure mediated by Ca2+-stimulated phospholipid scrambling was not different between red cells of the proband and control. Taken together, our findings imply that suppressed scrambling activity rather than flippase activity is the major contributor to maintainance of PS in inner leaflet of normal red cells during their 120-day lifespan, and that PS exposure to cell surface as an 'eat-me' signal depends primarily on scramblase activity at the end of lifespan. Importantly, our study has enables us to identify the major flippase of human erythrocyte membrane. Disclosures No relevant conflicts of interest to declare.

Sign in / Sign up

Export Citation Format

Share Document