Methods in Protein Sequence Analysis
Brigitte Wittmann-Liebold
- 01 Jan 1989
309
TL;DR: The AChR-binding sites on a-bungarotoxin by synthetic peptides representing the exposed regions of each of the toxin loops are localized by synthetic Peptide size selected to be within the optimum size requirement for T-cell activation.
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Abstract: A comprehensive synthetic approach, previously developed in this laboratory, has been applied to systematically screen the entire extracellular part (residues 1-210) of the a subunit of human and Torpedo californica acetylcholine receptors (AChR) for the various functional sites. Consecutive uniform-sized peptides were designed with constant 5-residue overlaps and covered the entire extracellular domain (residue 1-210) of I. californica and human AChR a subunits. The peptide size was selected to be within the optimum size requirement for T-cell activation. These pept i des were employed to 1 oca 1 i ze the cont i nuous regions recogni zed by anti-AChR antibodies, the regions recognized by AChR-primed T-lymphocytes and the regions that are involved in the binding of a-neurotoxins and of acetylcholine. We have also synthesized the two inter-transmembrane regions a262-276 and a-388408 as well as the f-terminal segment a427-437 and have determined their ability to bind a-neurotoxins (BgTX and CbTX) and antibodies against free AChR and membrane-bound AChR and to stimul ate the ~ vitro pro 1 i ferat i on of AChR-primed T-lymphocytes. This permitted the selection of a model for subunit organization within the membrane. In addition, we have also localized the AChR-binding sites on a-bungarotoxin by synthetic peptides representing the exposed regions of each of the toxin loops.
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TL;DR: The extrapolation of these findings to other known peptide antigens suggests that this tendency to form an amphipathic alpha-helix may be a general property of antigenic sites recognized by T cells, perhaps due to a different functional role of each type of site in eliciting T cell responses.
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Molecular cloning of cDNA coding for the gamma subunit of Torpedo acetylcholine receptor
TL;DR: This work describes a clone of 2,030 base pairs with sequences appropriate for the amino-terminal amino acids of the gamma subunit of acetylcholine receptor that hybridizes to a 2,100-base RNA present in electric organ but not in the brain of T. californica.
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S.D. Trakhanov,Marat Yusupov,Sultan Ch. Agalarov,Maria Garber,S.N. Ryazantsev,S.V. Tischenko,V.A. Shirokov +6 more
TL;DR: Well‐ordered three‐dimensional crystals from extremely thermophilic bacteria Thermus thermophilus reveal sedimentation constants of 70 S and 30 S, respectively, and are functionally active in the poly(U)‐system.
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