scholarly journals Haemoglobin binding with haptoglobin. Localization of the haptoglobin-binding site on the α-chain of human haemoglobin

1981 ◽  
Vol 197 (2) ◽  
pp. 507-510 ◽  
Author(s):  
A L Kazim ◽  
M Z Atassi
Keyword(s):  
Binding Site ◽  
Synthetic Approach ◽  
Protein Chain ◽  
Human Haemoglobin ◽  
Alpha Chain ◽  
Α Chain

A synthetic approach was employed to identify the haptoglobin-binding site on the alpha-chain of human haemoglobin. This approach cosists of the synthesis of a series of consecutive overlapping peptides that, together, systematically represent the entire protein chain. Fourteen peptides were synthesized (alpha 1-15, alpha 11-25, alpha 21-35, alpha 31-45, alpha 41-55, alpha 51-65, alpha 61-75, alpha 71-85, alpha 81-95, alpha 91-105, alpha 101-115, alpha 111-125, alpha 121-135 and alpha 131-141), and their ability bind human haptoglobin was studied, Only peptide alpha 121-135 bound haptoglobin significantly. On this basis we conclude that the haptoglobin-binding site on the alpha-chain of haemoglobin resides within, but does not necessarily encompass all of, the region alpha 121-135.

scholarly journals A novel and comprehensive synthetic approach for the elucidation of protein antigenic structures. Determination of the full antigenic profile of the α-chain of human haemoglobin

1980 ◽  
Vol 191 (1) ◽  
pp. 261-264 ◽  
Author(s):  
A L Kazim ◽  
M Z Atassi
Keyword(s):  
Primary Structure ◽  
Synthetic Approach ◽  
Human Haemoglobin ◽  
Alpha Chain ◽  
Antigenic Sites ◽  
Full Profile ◽  
Α Chain ◽  
First Time ◽  
Antigenic Profile

A comprehensive synthetic approach for the determination of continuous antigenic sites of proteins is presented. This approach consists of the synthesis of a series of consecutive overlapping peptides that, together, systematically represent the entire primary structure of the protein under study. Its application to the alpha-chain of human haemoglobin afforded, for the first time, a full profile of immunochemically active alpha-chain peptides and enabled the localization of all the major continuous antigenic sites of this haemoglobin subunit.

scholarly journals Structurally inherent antigenic sites. Localization of the antigenic sites of the α-chain of human haemoglobin in three host species by a comprehensive synthetic approach

1982 ◽  
Vol 203 (1) ◽  
pp. 201-208 ◽  
Author(s):  
A L Kazim ◽  
M Z Atassi
Keyword(s):  
Primary Structure ◽  
Host Species ◽  
Three Dimensional ◽  
Antigenic Structure ◽  
Synthetic Approach ◽  
Human Haemoglobin ◽  
Alpha Chain ◽  
Antigenic Sites ◽  
Full Profile ◽  
Related Proteins

The antigenic structure of the alpha-chain of human haemoglobin was studied by a synthetic approach consisting of the synthesis of a series of consecutive overlapping peptides that together systematically represent the entire primary structure of the protein. This approach enabled the identification of a full profile of immunochemically active alpha-chain peptides and the localization of its major ‘continuous’ antigenic sites. Antibodies to haemoglobin raised in each of three different species (goat, rabbit and mouse) recognize similar sites on the alpha-chain. Further, the molecular locations of these sites coincide with alpha-chain regions extrapolated from antigenic sites of the conformationally similar myoglobin molecule. These findings support our earlier proposed concept of ‘structurally inherent antigenic sites’, namely that antigenicity is conferred on certain surface regions of proteins by virtue of their three-dimensional locations. Thus the antigenic sites of conformationally related proteins are likely to have similar molecular locations.

scholarly journals Subunit interacting surfaces of human haemoglobin. Localization of the α-subunit-β-subunit interacting surfaces on the β-chain by a comprehensive synthetic strategy

1986 ◽  
Vol 234 (2) ◽  
pp. 457-461 ◽  
Author(s):  
N Yoshioka ◽  
M Z Atassi
Keyword(s):  
Binding Capacity ◽  
Binding Activity ◽  
The Other ◽  
Synthetic Approach ◽  
Beta Chain ◽  
Human Haemoglobin ◽  
Α Subunit ◽  
Alpha Chain ◽  
Synthetic Strategy ◽  
Interacting Surfaces

A synthetic approach is introduced for localization of subunit interacting surfaces in oligomeric proteins. It consists of studying the binding activity of consecutive uniform overlapping peptides encompassing an entire subunit to the other, radiolabelled, subunit. This permits the establishment of the full profile of peptides that bind the other intact subunit. This approach has been demonstrated with haemoglobin, and its application here with the beta-chain peptides has enabled the localization on the beta-chain of the submolecular regions responsible for its binding to alpha-chain in solution. There was good agreement between the binding surfaces found here in solution and those expected from the crystal structure. There were also, however, some significant differences in the levels of binding found in solution and those expected from the crystal. Peptide 21-35 possessed much higher binding activity than would be expected from its contribution to subunit association in the crystal. Conversely, other regions expected to possess considerable binding capacity for alpha-chain either showed low (peptides 111-125 and 121-135) or almost no binding (peptides 91-105 and 101-115) capacity. On the other hand, two interacting surfaces (within peptides 11-25 and 71-85) that make a contribution in solution do not appear to play a role in the crystal. It is concluded that the regions of subunit association in solution are close to, but not identical with, those in the crystal. The approach should serve as an effective method for localization of subunit interacting surfaces of unknown proteins, even those that can be isolated only in traces.

scholarly journals Haemoglobin binding with haptoglobin. Localization of the haptoglobin-binding sites on the β-chain of human haemoglobin by synthetic overlapping peptides encompassing the entire chain

1986 ◽  
Vol 234 (2) ◽  
pp. 453-456 ◽  
Author(s):  
N Yoshioka ◽  
M Z Atassi
Keyword(s):  
Binding Sites ◽  
Solid Phase ◽  
Three Dimensional ◽  
Binding Activity ◽  
Dimensional Structure ◽  
Synthetic Approach ◽  
Protein Chain ◽  
Beta Chain ◽  
Human Haemoglobin ◽  
Β Chain

A synthetic approach was employed to identify the haptoglobin-binding sites on the beta-chain of human haemoglobin. This approach consists of the synthesis of a series of consecutive overlapping peptides that, together, systematically represent the entire protein chain. Fourteen synthetic peptides (beta 1-15, beta 11-25 etc.) were examined for their ability to bind human haptoglobin by quantitative solid-phase radiometric titrations of 125I-labelled haptoglobin. Of these 14 peptides only peptides beta 11-25 and beta 131-146 bound haptoglobin significantly; peptide beta 21-35 exhibited a small binding activity as a consequence of the overlap with peptide beta 11-25. On this basis and by examination of the three-dimensional structure of haemoglobin, it was concluded that the beta-chain of haemoglobin has two binding sites for haptoglobin that reside in, but do not necessarily encompass all of, the regions beta 11-25 and beta 131-146.

scholarly journals Specific cyanylation and cleavage at cysteine-104 human hemoglobin α-chain. A novel approach to the problem of the α-chain tryptic core in the study of haemoglobin variants by ‘fingerprinting’ methods

1975 ◽  
Vol 145 (2) ◽  
pp. 251-261 ◽  
Author(s):  
R Casey ◽  
A Lang
Keyword(s):  
Gel Filtration ◽  
Peptide Bond ◽  
Complete Sequence ◽  
Cysteine Residue ◽  
Sequence Information ◽  
Human Haemoglobin ◽  
Alpha Chain ◽  
Novel Approach ◽  
Α Chain

1. A new approach to the analysis, by “fingerprinting”, of the tryptic core region of human haemoglobin alpha-chain is described. 2. The alpha-chain is cyanylated at its single cysteine residue (alpha104) and then split, by exposure to mild alkali, at the N-peptide bond of the resulting beta-thiocyanoalanine residue. 3. The two cleavage fragments, alpha1-103 and alpha104-141, are separated by gel filtration, and the fragment alpha104-141, which contains all the residues of the alpha-chain tryptic core, is digested with pepsin. 4. Preparative “fingerprints” of these peptic peptides yield eight major peptides, which provide complete sequence information for the whole region alpha104-141. 5. The utility of the method is demonstrated by repeating the determination of the substitution in haemoglobin Hopkins-2, a known alpha-chain core variant in which histidine-alpha112 (G19) is replaced by an aspartic acid residue.

Bipartite mechanism for laminin-integrin interactions: Identification of the integrin-binding site in LG domains of the laminin α chain

2020 ◽  
Vol 87 ◽  
pp. 66-76 ◽  
Author(s):  
Yukimasa Taniguchi ◽  
Mamoru Takizawa ◽  
Shaoliang Li ◽  
Kiyotoshi Sekiguchi
Keyword(s):  
Binding Site ◽  
Integrin Binding Site ◽  
Α Chain

scholarly journals Antigenic structure of human haemoglobin. Localization of the antigenic sites of the β-chain in three host species by synthetic overlapping peptides representing the entire chain

1986 ◽  
Vol 234 (2) ◽  
pp. 441-447 ◽  
Author(s):  
N Yoshioka ◽  
M Z Atassi
Keyword(s):  
Host Species ◽  
Structural Characteristics ◽  
Three Dimensional ◽  
Antigenic Structure ◽  
Synthetic Approach ◽  
Immune Recognition ◽  
Beta Chain ◽  
Human Haemoglobin ◽  
Antigenic Sites ◽  
Full Profile

A comprehensive synthetic approach is applied here to localize the continuous antigenic sites of the beta-chain of haemoglobin. The approach was based on the synthesis and purification of the following consecutive 15-residue peptides (each overlapping by five residues at both ends with the peptides preceding it and following it in the sequence): 1-15, 11-25 etc. Quantitative radiometric titrations of protein and peptide adsorbents were performed with 125I-labelled anti-haemoglobin antibodies from three different host species. The specificity of antibody binding to peptide adsorbents was confirmed by inhibition studies and by the binding specificity of antibodies isolated from peptide adsorbents. These studies established the full profile of antigenic beta-chain regions, which was found to be independent of the host species. Five major antigenic sites were localized, and their three-dimensional and structural characteristics are discussed in relation to the immune recognition of haemoglobin and other proteins.

scholarly journals Mapping of the properdin-binding site in the third component of complement

1984 ◽  
Vol 217 (1) ◽  
pp. 323-326 ◽  
Author(s):  
J D Lambris ◽  
J Alsenz ◽  
T F Schulz ◽  
M P Dierich
Keyword(s):  
Binding Site ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Enzyme Linked Immunosorbent Assay ◽  
Chain Fragment ◽  
Alpha Chain ◽  
Factor I ◽  
Assay Procedure ◽  
Complement Component C3 ◽  
Third Component Of Complement

The properdin-binding site in the human third complement component (C3) was mapped by using isolated C3b, C3c, alpha- and beta-chains of C3 and C3 polypeptide fragments and an enzyme-linked-immunosorbent-assay procedure. The C3 chains and the polypeptide fragments were purified to homogeneity by preparative sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. The alpha-chain polypeptides included a 68 kDa and a 43 kDa polypeptide, which were generated by cleavage of C3b with factors I and H, and a 40 kDa, 33 kDa (C3d) and 27 kDa polypeptide, which were generated by cleavage of C3b with porcine elastase. It was shown that properdin binds to C3b, C3c, alpha-chain, and to the 43 kDa (factor-I + H-derived), as well as to 40 kDa (elastase-derived) alpha-chain fragment, but not to the beta-chain 68 kDa, 33 kDa (C3d) and 27 kDa alpha-chain fragments. Thus the binding site for properdin resides on the 40-43 kDa C-terminal alpha-chain fragment of C3.

scholarly journals Ciliary Neurotrophic Factor, Cardiotrophin-like Cytokine, and Neuropoietin Share a Conserved Binding Site on the Ciliary Neurotrophic Factor Receptor α Chain

2008 ◽  
Vol 283 (44) ◽  
pp. 30341-30350 ◽  
Author(s):  
François Rousseau ◽  
Sylvie Chevalier ◽  
Catherine Guillet ◽  
Elisa Ravon ◽  
Caroline Diveu ◽  
...  
Keyword(s):  
Binding Site ◽  
Neurotrophic Factor ◽  
Ciliary Neurotrophic Factor ◽  
Α Chain ◽  
Ciliary Neurotrophic Factor Receptor
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