scholarly journals Localization of the protein 4.1-binding site on human erythrocyte glycophorins C and D

1994 ◽  
Vol 299 (1) ◽  
pp. 191-196 ◽  
Author(s):  
N J Hemming ◽  
D J Anstee ◽  
W J Mawby ◽  
M E Reid ◽  
M J Tanner
Keyword(s):  
Amino Acid ◽  
Human Erythrocyte ◽  
Synthetic Peptide ◽  
Attachment Site ◽  
Amino Acid Residues ◽  
Protein 4.1 ◽  
Attachment Sites ◽  
Underlying Network ◽  
Membrane Attachment ◽  
Glycophorin C

The flexibility of the human erythrocyte membrane is mediated by an underlying network of skeletal proteins which interact with the membrane through ankyrin and protein 4.1. The nature of the membrane attachment site(s) for protein 4.1 has yet to be fully elucidated. In this paper we show that purified protein 4.1 binds much less strongly to alkali-stripped membranes from erythrocytes of individuals with total glycophorin C and D deficiency (Leach phenotype) than to alkali-stripped normal membranes. We further show that a synthetic peptide corresponding to amino acid residues 82-98 of the cytoplasmic domain of glycophorin C specifically binds to purified protein 4.1 and inhibits protein 4.1 binding to alkali-stripped normal membranes. The same synthetic peptide binds directly to membranes from individuals with glycophorin C and D deficiency but not to normal membranes. These results indicate that glycophorins C and D provide major membrane attachment sites for protein 4.1 in normal erythrocytes and that this interaction is mediated by protein 4.1 binding to amino acid residues 82-98 of glycophorin C and 61-77 of glycophorin D.

scholarly journals Quantitation of the number of molecules of glycophorins C and D on normal red blood cells using radioiodinated Fab fragments of monoclonal antibodies

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1668-1672 ◽  
Author(s):  
J Smythe ◽  
B Gardner ◽  
DJ Anstee
Keyword(s):  
Monoclonal Antibodies ◽  
Blood Cells ◽  
Binding Assays ◽  
Membrane Glycoproteins ◽  
Human Erythrocyte Membrane ◽  
Protein 4.1 ◽  
Fab Fragments ◽  
Attachment Sites ◽  
Membrane Attachment ◽  
Glycophorin C

Abstract Two rat monoclonal antibodies (BRAC 1 and BRAC 11) have been produced. BRAC 1 recognizes an epitope common to the human erythrocyte membrane glycoproteins glycophorin C (GPC) and glycophorin D (GPD). BRAC 11 is specific for GPC. Fab fragments of these antibodies and BRIC 10, a murine monoclonal anti-GPC, were radioiodinated and used in quantitative binding assays to measure the number of GPC and GPD molecules on normal erythrocytes. Fab fragments of BRAC 11 and BRIC 10 gave values of 143,000 molecules GPC per red blood cell (RBC). Fab fragments of BRAC 1 gave 225,000 molecules of GPC and GPD per RBC. These results indicate that GPC and GPD together are sufficiently abundant to provide membrane attachment sites for all of the protein 4.1 in normal RBCs.

scholarly journals Quantitation of the number of molecules of glycophorins C and D on normal red blood cells using radioiodinated Fab fragments of monoclonal antibodies

Blood ◽  
1994 ◽  
Vol 83 (6) ◽  
pp. 1668-1672
Author(s):  
J Smythe ◽  
B Gardner ◽  
DJ Anstee
Keyword(s):  
Monoclonal Antibodies ◽  
Blood Cells ◽  
Binding Assays ◽  
Membrane Glycoproteins ◽  
Human Erythrocyte Membrane ◽  
Protein 4.1 ◽  
Fab Fragments ◽  
Attachment Sites ◽  
Membrane Attachment ◽  
Glycophorin C

Two rat monoclonal antibodies (BRAC 1 and BRAC 11) have been produced. BRAC 1 recognizes an epitope common to the human erythrocyte membrane glycoproteins glycophorin C (GPC) and glycophorin D (GPD). BRAC 11 is specific for GPC. Fab fragments of these antibodies and BRIC 10, a murine monoclonal anti-GPC, were radioiodinated and used in quantitative binding assays to measure the number of GPC and GPD molecules on normal erythrocytes. Fab fragments of BRAC 11 and BRIC 10 gave values of 143,000 molecules GPC per red blood cell (RBC). Fab fragments of BRAC 1 gave 225,000 molecules of GPC and GPD per RBC. These results indicate that GPC and GPD together are sufficiently abundant to provide membrane attachment sites for all of the protein 4.1 in normal RBCs.

scholarly journals Identification of the Membrane Attachment Sites for Protein 4.1 in the Human Erythrocyte

1995 ◽  
Vol 270 (10) ◽  
pp. 5360-5366 ◽  
Author(s):  
Nicola J. Hemming ◽  
David J. Anstee ◽  
Marcelo A. Staricoff ◽  
Michael J. A. Tanner ◽  
Narla Mohandas
Keyword(s):  
Human Erythrocyte ◽  
Protein 4.1 ◽  
Attachment Sites ◽  
Membrane Attachment

Plasmin-platelet interaction involves cleavage of functional thrombin receptor

1996 ◽  
Vol 271 (1) ◽  
pp. C54-C60 ◽  
Author(s):  
M. Kimura ◽  
T. T. Andersen ◽  
J. W. Fenton ◽  
W. F. Bahou ◽  
A. Aviv
Keyword(s):  
Amino Acid ◽  
Amino Acid Sequence ◽  
Platelet Activation ◽  
Synthetic Peptide ◽  
Time Dependent ◽  
Thrombin Receptor ◽  
Amino Acid Residues ◽  
Enzymatic Action ◽  
High Concentrations ◽  
Inhibition Of Thrombin

We tested the hypothesis that the inhibition of thrombin-induced platelet activation by plasmin is mediated via the enzymatic action of plasmin on the functional thrombin receptor. We monitored the binding of the anti-thrombin receptor antibody [anti-TR-(34-46)] to platelets; this binding is sensitive to the cleavage of the thrombin receptor at amino acid residues Arg-41 to Ser-42. Plasmin inhibited anti-TR-(34-46) binding in dose- and time-dependent manners. The inactive synthetic peptide with the amino acid sequence 40-55 of the thrombin receptor (D-FPRSFLLRNPNDKYEPF) was similarly cleaved by thrombin and plasmin to an active peptide (SFLLRNPNDKYEPF) that produced robust cytosolic Ca2+ responses. At high concentrations, plasmin itself can activate platelets. We explored this effect with the use of anti-TR-(1-160). This antibody abolished the cytosolic Ca2+ responses to thrombin and to the thrombin receptor-activating peptide SFLLRN but did not attenuate the plasmin-induced cytosolic Ca2+ response. Thus plasmin inhibits thrombin-evoked platelet activation by cleaving the thrombin receptor, but the plasmin-induced cytosolic Ca2+ response is not due to the generation of the tethered peptide of the thrombin receptor.

Nucleotide sequence of cDNA encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP3

1995 ◽  
Vol 7 (5) ◽  
pp. 1209 ◽  
Author(s):  
SK Kolluri ◽  
R Kaul ◽  
K Banerjee ◽  
SK Gupta
Keyword(s):  
Amino Acid ◽  
Zona Pellucida ◽  
Amino Acid Residues ◽  
Reading Frame ◽  
Contraceptive Vaccine ◽  
Attachment Sites ◽  
Bonnet Monkey ◽  
Polymerase Chain ◽  
Human Use ◽  
Sugar Chains

The cDNA encoding bonnet monkey zona pellucida ZP3 from bonnet ovary has been amplified by polymerase chain reaction. The ZP3 gene has an open reading frame of 1272 nucleotides encoding a polypeptide of 424 amino acid residues which shares 93.9% overall identity with human ZP3. Bonnet ZP3 has four potential attachment sites for N-linked sugar chains which are also conserved in human ZP3. Bonnet ZP3 has 14 cysteine residues compared with 15 in human ZP3. The highest disparity between these molecules was restricted to a domain represented by amino acid residues 370-398. These results have important implications for the use of bonnet monkey as an animal model for evaluation and development of contraceptive vaccine based on ZP3 for human use.

The human anion transport protein, band 3, contains a CD36-like binding domain forPlasmodium falciparum-infected erythrocytes

Parasitology ◽  
1996 ◽  
Vol 112 (3) ◽  
pp. 261-267 ◽  
Author(s):  
I. Crandall ◽  
I. W. Sherman
Keyword(s):  
Monoclonal Antibody ◽  
Amino Acid ◽  
Blood Cells ◽  
Synthetic Peptide ◽  
Epitope Mapping ◽  
Protein Band ◽  
Band 3 ◽  
Anion Transport ◽  
Transport Protein ◽  
Amino Acid Residues

SUMMARYEpitope mapping of a murine monoclonal antibody (mAb), 5H12, prepared against livePlasmodium falciparum-intected red blood cells indicated that the epitope consisted of amino acid residues 474–487 of the human anion transport protein, band 3. mAb 5H12 enhanced cytoadherence, but inhibited the CD36-like mediated resetting. A synthetic peptide based on the sequence of the epitope (FSFCETNGLE) blocked both resetting and cytoadherence, suggesting that this amino acid sequence may form the CD36-like receptor. The CD36-like region of band 3 was antigenically distinct from platelet or endothelial CD36.

Immunization with human thyrotrophin receptor peptide induces an increase in thyroid hormone in rabbits

1992 ◽  
Vol 135 (3) ◽  
pp. 479-484 ◽  
Author(s):  
M. Ohmori ◽  
T. Endo ◽  
M. Ikeda ◽  
T. Onaya
Keyword(s):  
Amino Acid ◽  
Thyroid Hormone ◽  
Western Blot ◽  
Synthetic Peptide ◽  
Matched Control ◽  
Tsh Receptor ◽  
Amino Acid Residues ◽  
Normal Range ◽  
Tsh Receptor Antibody ◽  
Peptide Antibodies

ABSTRACT Eight rabbits were immunized with a synthetic peptide corresponding to the unique N-terminal region (termed N peptide; amino acid residues 29–57) in the extracellular domain of the human thyrotrophin (TSH) receptor. After 10 weeks, all of the eight rabbits produced anti-N peptide antibodies. Western blot analysis revealed that the antibodies recognized rabbit TSH receptor as an approximately 100 kDa protein. We compared the level of thyroid hormone in serum taken before immunization (preimmune sera) with that of serum taken after immunization (postimmune sera) in these immunized rabbits. Postimmune sera from the eight rabbits had higher mean (± s.d.) levels of tri-iodothyronine (T3) and thyroxine (T4) than did preimmune sera (T3, preimmune 0·82 ± 0·26 μg/l vs postimmune 1·33 ± 0·35, P < 0·01; T4, preimmune 33·7 ± 10·0 μg/l vs postimmune 41·0 ± 6·0, P < 0·05). T3 levels in four rabbits and T4 levels in four rabbits after immunization were over the normal range obtained from six age-matched control rabbits. Seven rabbits exhibited thyroid-stimulating antibody (TSAb) activity with various degrees (241–545%). The concentration of T3 and T4 did not increase over 10 weeks in either non-immunized rabbits (T3, preimmune 0·89 ± 0·34 μg/l vs postimmune 0·82 ± 0·22; T4, preimmune 31·1 ± 7·3 μg/l vs postimmune 30·3 ± 5·1) or other peptide-immunized rabbits (T3, preimmune 0·68 μg/l (n = 2) vs postimmune 0·69; T4, preimmune 33·1 μg/l vs postimmune 26·4). These results indicate that experimentally produced anti-TSH receptor antibody with TSAb activity induces an increase in thyroid hormone in rabbits. Journal of Endocrinology (1992) 135, 479–484

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