Evidence for an Asymmetric Distribution of Phospholipids in the Human Erythrocyte Membrane

1974 ◽  
Vol 52 (9) ◽  
pp. 803-806 ◽  
Author(s):  
Arthur Kahlenberg ◽  
Caroline Walker ◽  
Ruth Rohrlick
Keyword(s):  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Phospholipid Composition ◽  
Enzyme Hydrolysis ◽  
Asymmetric Distribution ◽  
Human Erythrocyte Membrane ◽  
Cytoplasmic Surface ◽  
Direct Demonstration ◽  
Inside Out ◽  
Phospholipases A

The changes in phospholipid composition of the inner (cytoplasmic) surface of the human erythrocyte membrane resulting from the digestion of sealed inside-out vesicles with phospholipases A2 and C were determined. Virtually all of the phosphatidylethanolamine and phosphatidylserine and 30–40% of the phosphatidylcholine and sphingomyelin of inside-out vesicles were found to be accessible to enzyme hydrolysis. In contrast, all of the above phospholipids of unsealed ghosts were susceptible to phospholipolytic digestion. These results are a direct demonstration of an asymmetric distribution of phospholipids in the human erythrocyte membrane.

scholarly journals Appearance and distribution of surface proteins of the human erythrocyte membrane. An electron microscope and immunochemical labeling study.

1978 ◽  
Vol 76 (2) ◽  
pp. 512-531 ◽  
Author(s):  
D Shotton ◽  
K Thompson ◽  
L Wofsy ◽  
D Branton
Keyword(s):  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Indirect Evidence ◽  
Surface Proteins ◽  
Human Erythrocyte Membrane ◽  
Intramembrane Particle ◽  
Translational Movement ◽  
Cytoplasmic Surface ◽  
Particle Aggregates ◽  
Before And After

We have used freeze-etching, before and after immunoferritin labeling, to visualize spectrin molecules and other surface proteins of the human erythrocyte membrane. After intramembrane particle aggregation was induced, spectrin molecules, identified by labeling with ferritin-conjugated antispectrin, were clustered on the cytoplasmic surface of the membrane in patches directly underlying the particle clusters. This labeling pattern confirms the involvement of spectrin in such particle aggregates, as previously inferred from indirect evidence. Ferritin-conjugated antihapten molecules, directed against external and cytoplasmic surface proteins of the erythrocyte membrane which had been covalently labeled nonspecifically with the hapten p-diazoniumphenyl-beta-D-lactoside, were similarly found in direct association with such intramembrane particle aggregates. This indicates that when spectrin and the intramembrane particles are aggregated, all the major proteins of the erythrocyte membrane are constrained to coaggregate with them. Although giving no direct information concerning the freedom of translational movement of proteins in the unperturbed erythrocyte membrane, these experiments suggest that a close dynamic association may exist between the integral and peripheral protein components of the membrane, such that immobilization of one component can restrict the lateral mobility of others.

Asymmetric distribution of phosphoinositides and phosphatidic acid in the human erythrocyte membrane

1991 ◽  
Vol 1069 (1) ◽  
pp. 27-36 ◽  
Author(s):  
Philippe Gascard ◽  
Dien Tran ◽  
Monique Sauvage ◽  
Jean-Claude Sulpice ◽  
Kiyoko Fukami ◽  
...  
Keyword(s):  
Phosphatidic Acid ◽  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Asymmetric Distribution ◽  
Human Erythrocyte Membrane

scholarly journals Electron microscope study of reassociation of spectrin and actin with the human erythrocyte membrane

1981 ◽  
Vol 90 (1) ◽  
pp. 70-77 ◽  
Author(s):  
S Tsukita ◽  
S Tsukita ◽  
H Ishikawa ◽  
S Sato ◽  
M Nakao
Keyword(s):  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Actin Filaments ◽  
Actin Polymerization ◽  
Erythrocyte Membranes ◽  
Human Erythrocyte Membrane ◽  
Thin Sections ◽  
Fixation Treatment ◽  
Human Erythrocyte Membranes ◽  
Inside Out

Reassociation of spectrin and actin with human erythrocyte membranes was studied by stereoscopic electron microscopy of thin sections combined with tannic acid- glutaraldehyde fixation. Treatment of the erythrocyte membrane with 0.1 mM EDTA (pH 8.0) extracted more than 90 percent of the spectrin and actin and concomitantly removed filamentous meshworks underlying the membranes, followed by fragmentation into small inside-out vesicles. When such spectrin-depleted vesicles were incubated with the EDTA extract (crude spectrin), a filamentous meshwork, similar to those of the original membranes, was reformed on the cytoplasmic surface of the vesicles. The filamentous components, with a uniform thickness of 9 nm, took a tortuous course and joined one another often in an end-to-end fashion to form a irregular but continuous meshwork parallel to the membrane. Purified spectrin was also reassociated with the vesicles in a population density of filamentous components almost comparable to that of the crude spectrin-reassociated vesicles. However, the meshwork formation was much smaller in extent, showing many independent filamentous components closely applied to the vesicle surface. When muscle G-actin was added to the crude spectrin- or purified spectrin- reassociated vesicles under conditions which favor actin polymerization, actin filaments were seen to attach to the vesicles through the filamentous components. Two modes of association of actin filaments with the membrane were seen: end-to-membrane and side-to- membrane associations. In the end-to-membrane association, each actin filament was bound with several filamentous components exhibiting a spiderlike configuration, which was considered to be the unit of the filamentous meshwork of the original erythrocyte membrane.

scholarly journals Purification of the NF2 Tumor Suppressor Protein from Human Erythrocytes

2006 ◽  
Vol 33 (4) ◽  
pp. 394-402 ◽  
Author(s):  
Hitesh K. Jindal ◽  
Kazumi Yoshinaga ◽  
Pil-Soo Seo ◽  
Mohini Lutchman ◽  
Patrick A. Dion ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Tumor Suppressor ◽  
Erythrocyte Membrane ◽  
Human Erythrocyte ◽  
Mammalian Cells ◽  
Tumor Suppressor Protein ◽  
Human Erythrocyte Membrane ◽  
Protein 4.1 ◽  
Erythrocyte Plasma Membrane ◽  
Inside Out

Background:Neurofibromatosis type 2 (NF2) is an autosomal dominant disease predisposing individuals to the risk of developing tumors of cranial and spinal nerves. The NF2 tumor suppressor protein, known as Merlin/Schwanomin, is a member of the protein 4.1 superfamily that function as links between the cytoskeleton and the plasma membrane.Methods:Upon selective extraction of membrane-associated proteins from erythrocyte plasma membrane (ghosts) using low ionic strength solution, the bulk of NF2 protein remains associated with the spectrin-actin depleted inside-out-vesicles. Western blot analysis showed a ~70 kDa polypeptide in the erythrocyte plasma membrane. Furthermore, quantitative removal of NF2 protein from the inside-out-vesicles was achieved using 1.0 M potassium iodide, a treatment known to remove tightly-bound peripheral membrane proteins.Results:These results suggest a novel mode of NF2 protein association with the erythrocyte membrane that is distinct from the known membrane interactions of protein 4.1. Based on these biochemical properties, several purification strategies were devised to isolate native NF2 protein from human erythrocyte ghosts. Using purified and recombinant NF2 protein as internal standards, we quantified approximately ~41-65,000 molecules of NF2 protein per erythrocyte.Conclusion:We provide evidence for the presence of NF2 protein in the human erythrocyte membrane. The identification of NF2 protein in the human erythrocyte membrane will make it feasible to discover novel interactions of NF2 protein utilizing powerful techniques of erythrocyte biochemistry and genetics in mammalian cells.

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