scholarly journals Monoclonal antibodies to the membrane domain of the human erythrocyte anion transport protein. Localization of the C-terminus of the protein to the cytoplasmic side of the red cell membrane and distribution of the protein in some human tissues

1989 ◽  
Vol 258 (1) ◽  
pp. 211-220 ◽  
Author(s):  
S D Wainwright ◽  
M J A Tanner ◽  
G E M Martin ◽  
J E Yendle ◽  
C Holmes
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Membrane ◽  
Human Erythrocyte ◽  
Anion Transport ◽  
Transport Protein ◽  
Red Cell ◽  
Membrane Domain ◽  
Red Cell Membrane ◽  
C Terminus ◽  
Cytoplasmic Side

(1) We have prepared murine monoclonal antibodies to the membrane domain of the human erythrocyte anion transport protein (band 3). (2) All of these antibodies react with regions of the protein located at the cytoplasmic surface of the red cell. (3) One of the antibodies reacts with an epitope present on a cytoplasmic loop of the protein located between the C-terminus and a point 168 amino acids from the C-terminus. The other antibodies recognize different epitopes on the C-terminal tail of the protein and the sequences likely to be involved in these epitopes are defined. (4) Our results show that the C-terminus of the red-cell anion transport protein is located on the cytoplasmic side of the red-cell membrane. (5) None of the antibodies inhibited sulphate exchange transport when introduced into resealed red-cell membranes; however, the bivalent form of one of the antibodies reduced the inhibitory potency of 4-acetamido-4'-isothiocyanatostilbene disulphonate on sulphate exchange transport in resealed erythrocyte membranes. (6) Immunostaining of human kidney sections with the antibodies showed strong staining of the basolateral membrane of some but not all of the epithelial cells of distal tubules and the initial connecting segment of collecting tubules. With human liver, only the haematopoeitic cells of fetal liver reacted with all the antibodies.

scholarly journals The membrane domain of the human erythrocyte anion transport protein. Epitope mapping of a monoclonal antibody defines the location of a cytoplasmic loop near the C-terminus of the protein

1990 ◽  
Vol 272 (1) ◽  
pp. 265-268 ◽  
Author(s):  
S D Wainwright ◽  
W J Mawby ◽  
M J A Tanner
Keyword(s):  
Amino Acid ◽  
Human Erythrocyte ◽  
Protein Band ◽  
Anion Transport ◽  
Transport Protein ◽  
Amino Acid Sequences ◽  
Amino Acid Residues ◽  
C Terminus ◽  
Murine Monoclonal Antibodies ◽  
Cytoplasmic Side

We have used synthetic peptides to study the location of the amino acid sequences in the human erythrocyte anion transport protein (band 3) which are recognized by four murine monoclonal antibodies, BRIC 130, 132, 154 and 155. These antibodies are known to react with epitopes in the protein which are on the cytoplasmic side of the membrane. The results suggest that the amino acid residues important for the reaction of BRIC 130 and BRIC 154/155 are located within amino acids 899-908 and 895-901 respectively in the cytoplasmic tail of the protein. The BRIC 132 epitope is located within amino acid residues 813-824. This is part of a surface loop in the protein which probably extends from residue 814 to residue 832 and is located on the cytoplasmic side of the membrane. These results provide direct evidence for the topographical location of a sequence in a poorly understood region of the protein.

scholarly journals The human erythrocyte anion-transport protein. Further amino acid sequence from the integral membrane domain homologous with the murine protein

1986 ◽  
Vol 235 (3) ◽  
pp. 899-901 ◽  
Author(s):  
C J Brock ◽  
M J A Tanner
Keyword(s):  
Amino Acid ◽  
Human Erythrocyte ◽  
Proteolytic Enzymes ◽  
Transmembrane Helix ◽  
Anion Transport ◽  
Transport Protein ◽  
Membrane Domain ◽  
C Terminus ◽  
N Terminus ◽  
Almost All

Sequences from the human erythrocyte anion-transport protein homologous with residues 417-449 and 794-813 of the murine erythrocyte anion-transport protein have been determined. The former sequence includes the putative transmembrane helix closest to the N-terminus of the protein. The latter sequence traverses almost all of the lipid bilayer and is located towards the C-terminus of the protein. Sites have been identified by alignment with the murine sequence in the integral membrane domain that are accessible to proteolytic enzymes. Sequences from the integral membrane domain of the erythrocyte anion-transport protein are highly conserved.

scholarly journals Inhibition of malarial parasite invasion by monoclonal antibodies against glycophorin A correlates with reduction in red cell membrane deformability

Blood ◽  
1989 ◽  
Vol 74 (5) ◽  
pp. 1836-1843 ◽  
Author(s):  
G Pasvol ◽  
JA Chasis ◽  
N Mohandas ◽  
DJ Anstee ◽  
MJ Tanner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Membrane ◽  
Red Cells ◽  
Malarial Parasite ◽  
Glycophorin A ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Parasite Invasion ◽  
Surface Molecules ◽  
Fab Fragments

Abstract The effect of well-characterized monoclonal antibodies to red cell surface molecules on the invasion of human red cells by the malarial parasites Plasmodium falciparum and Plasmodium knowlesi was examined. Antibodies to glycophorin A (GP alpha) inhibit invasion for both parasite species, and this is highly correlated with the degree to which they decrease red cell membrane deformability as measured by ektacytometry. This effect on rigidity and invasion was also seen with monovalent Fab fragments. The closer the antibody binding site was to the membrane bilayer, the greater was its effect on inducing membrane rigidity and decreasing parasite invasion. Antibodies to the Wright determinant in particular were the most inhibitory. This differential effect of the various antibodies was not correlated with their binding affinities or the number of sites bound per cell. Antibodies to surface molecules other than GP alpha were without effect. A novel mechanism is described whereby monoclonal antibodies and their Fab fragments directed at determinants on the external surface of red cells might act to inhibit invasion by malarial parasites by altering membrane material properties.

scholarly journals The complete amino acid sequence of the human erythrocyte membrane anion-transport protein deduced from the cDNA sequence

1988 ◽  
Vol 256 (3) ◽  
pp. 703-712 ◽  
Author(s):  
M J A Tanner ◽  
P G Martin ◽  
S High
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Protein Band ◽  
Anion Transport ◽  
Transport Protein ◽  
Cdna Clones ◽  
Red Cell ◽  
Coding Region ◽  
Red Cell Membrane ◽  
Anion Transporter

1. We have isolated cDNA clones corresponding to the red cell membrane anion-transport protein (Band 3). 2. The cDNA clones cover 3475 bases of the mRNA and contain the entire protein-coding region, 150 bases of the 5′ untranslated region and part of the 3′ non-coding region, but do not extend to the 3′ end of the mRNA. 3. The translated protein sequence predicts that the human red cell anion transporter contains 911 amino acids. 4. The availability of the amino acid sequence allows the interpretation of some of the many studies on the chemical and proteolytic modification of the human protein aimed at examining the structure and mechanism of this membrane transport protein.

Inhibition of malarial parasite invasion by monoclonal antibodies against glycophorin A correlates with reduction in red cell membrane deformability

Blood ◽  
1989 ◽  
Vol 74 (5) ◽  
pp. 1836-1843
Author(s):  
G Pasvol ◽  
JA Chasis ◽  
N Mohandas ◽  
DJ Anstee ◽  
MJ Tanner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Membrane ◽  
Red Cells ◽  
Malarial Parasite ◽  
Glycophorin A ◽  
Red Cell ◽  
Red Cell Membrane ◽  
Parasite Invasion ◽  
Surface Molecules ◽  
Fab Fragments

The effect of well-characterized monoclonal antibodies to red cell surface molecules on the invasion of human red cells by the malarial parasites Plasmodium falciparum and Plasmodium knowlesi was examined. Antibodies to glycophorin A (GP alpha) inhibit invasion for both parasite species, and this is highly correlated with the degree to which they decrease red cell membrane deformability as measured by ektacytometry. This effect on rigidity and invasion was also seen with monovalent Fab fragments. The closer the antibody binding site was to the membrane bilayer, the greater was its effect on inducing membrane rigidity and decreasing parasite invasion. Antibodies to the Wright determinant in particular were the most inhibitory. This differential effect of the various antibodies was not correlated with their binding affinities or the number of sites bound per cell. Antibodies to surface molecules other than GP alpha were without effect. A novel mechanism is described whereby monoclonal antibodies and their Fab fragments directed at determinants on the external surface of red cells might act to inhibit invasion by malarial parasites by altering membrane material properties.

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