Analysis of Receptor for Vibrio choleraeEl Tor Hemolysin with a Monoclonal Antibody That Recognizes Glycophorin B of Human Erythrocyte Membrane

ABSTRACT El Tor hemolysin (ETH), a pore-forming toxin secreted byVibrio cholerae O1 biotype El Tor and most Vibrio cholerae non-O1 isolates, is able to lyse erythrocytes and other mammalian cells. To study the receptor for this toxin or the related molecule(s) on erythrocyte, we first isolated a monoclonal antibody, B1, against human erythrocyte membrane, which not only blocks the binding of ETH to human erythrocyte but also inhibits the hemolytic activity of ETH. Biochemical characterization and immunoblotting revealed that this antibody recognized an epitope on the extracellular domain of glycophorin B, a sialoglycoprotein of erythrocyte membrane. Erythrocytes lacking glycophorin B but not glycophorin A were less sensitive to the toxin than were normal human erythrocytes. These results indicate that glycophorin B is a receptor for ETH or at least an associated molecule of the receptor for ETH on human erythrocytes.

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Degradation of the human erythrocyte membrane band 3 studied with the monoclonal antibody directed against an epitope on the cytoplasmic fragment of band 3

European Journal of Biochemistry ◽

10.1111/j.1432-1033.1988.tb14147.x ◽

1988 ◽

Vol 174 (4) ◽

pp. 647-654 ◽

Cited By ~ 32

Author(s):

Marcin CZERWINSKI ◽

Kazimiera WASNIOWSKA ◽

Iga STEUDEN ◽

Maria DUK ◽

Antoni WIEDLHA ◽

...

Keyword(s):

Monoclonal Antibody ◽

Erythrocyte Membrane ◽

Human Erythrocyte ◽

Band 3 ◽

Human Erythrocyte Membrane

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Purification of the NF2 Tumor Suppressor Protein from Human Erythrocytes

Canadian Journal of Neurological Sciences / Journal Canadien des Sciences Neurologiques ◽

10.1017/s0317167100005357 ◽

2006 ◽

Vol 33 (4) ◽

pp. 394-402 ◽

Cited By ~ 2

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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Uptake and binding of disaccharides in human erythrocytes

Canadian Journal of Biochemistry ◽

10.1139/o76-016 ◽

1976 ◽

Vol 54 (1) ◽

pp. 99-101 ◽

Cited By ~ 2

Author(s):

Ivan Beneš ◽

Arnošt Kotyk

Keyword(s):

Erythrocyte Membrane ◽

Human Erythrocyte ◽

Intact Cell ◽

Binding Capacity ◽

Human Erythrocytes ◽

Human Erythrocyte Membrane ◽

Maximum Binding Capacity

Disaccharides (sucrose, lactose, melibiose, cellobiose, trehalose, maltose, and isomaltose) are not transported across the human erythrocyte membrane. Maltose alone is bound in appreciable amounts to the intact cell as well as ghost membranes and competes mutually for uptake with D-glucose. In (NH4)2SO4-precipitated membrane preparations, maltose binds more strongly than other disaccharides (KD = 1.3 × 10−5 M; maximum binding capacity, 71 pmol/mg protein) and again competes mutually with D-glucose. Phloretin inhibits the binding of glucose much more than that of maltose.

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Facilitated transport of amino acids across organic phases and the human erythrocyte membrane

Biochemical Journal ◽

10.1042/bj1880541 ◽

1980 ◽

Vol 188 (2) ◽

pp. 541-548 ◽

Cited By ~ 6

Author(s):

R C Hider ◽

W McCormack

Keyword(s):

Amino Acids ◽

Amino Acid ◽

Erythrocyte Membrane ◽

Transfer Process ◽

Human Erythrocyte ◽

Facilitated Transport ◽

Human Erythrocytes ◽

Human Erythrocyte Membrane ◽

Chloroform Phase

1. An artificial facilitated amino-acid-transfer process operating across a chloroform phase is reported. 2. This process utilizes a family of bis(salicylamidato)copper(II) complexes. 3. A mechanism is proposed for this process and for its sensitivity towards cyanide and bathophenanthroline sulphonate. 4. Facilitated transfer of L-leucine in human erythrocytes has been shown to be inhibited by bathophenanthroline sulphonate.

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