Strategies for epitope analysis using peptide synthesis

Geysen, H. Mario; Rodda, Stuart J.; Mason, Tom J.; Tribbick, Gordon; Schoofs, Peter

doi:10.1016/0022-1759(87)90085-8

Cited by 853 publications

(480 citation statements)

References 25 publications

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“…Similar data were obtained by Stephen et al (1995) who reported that moAbs, including antip53 moAb, with known peptide reactivity, selected from 12-mer and 20-mer phage-displayed libraries not only the expected peptides but also sequences with no discernible homology with the original antigen. Presence of epitopes de®ned as common antigenic determinants and characterized by no primary sequence homology has been previously mentioned and called`mimotopes' by Geysen et al, 1986Geysen et al, , 1987. Interaction of a speci®c moAb with both epitope and mimotope motifs was explained by interaction with the two di erent ligands either of distinct paratopes on the same moAb or alternative contacts of the same paratope (Stephen et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

Identification of RB18A, a 205 kDa new p53 regulatory protein which shares antigenic and functional properties with p53

et al. 1997

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Immunological screening with the anti-p53 moAb, PAb1801 of a cDNA expression library, prepared from human B lymphoma cells, led us to identify a new human 205 kDa protein called RB18A for`Recognized By PAb1801 moAntibody'. Immunoblotting or immunoprecipitation of fusion protein or in vitro translated protein, respectively, demonstrated that RB18A protein was recognized by several anti-p53 moAb reacting with the N or C-terminal domains of p53. Full length sequence of RB18A cDNA and computer analysis demonstrated that despite common antigenic determinants between RB18A and p53 proteins, nucleotide and deduced protein sequences did not reveal any signi®cant homologies. RB18A mRNA was detected in all tissues tested except in kidney. In addition, RB18A protein shared identical functions with p53 protein: binding to DNA or to p53 and self-oligomerization. Furthermore, RB18A regulated p53 speci®c binding on his DNA consensus binding site. These functions were associated to the C-terminal domain of RB18A protein and more speci®cally to the PAb421 binding site present in this domain. The activation by RB18A of p53 binding on DNA was induced through an unstable interaction between both proteins. Altogether, our data demonstrated that RB18A protein shares antigenic and functional properties with p53 and regulated p53 functions.

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Section: Discussionmentioning

confidence: 99%

Identification of RB18A, a 205 kDa new p53 regulatory protein which shares antigenic and functional properties with p53

et al. 1997

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“…Antibodies were first tested by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting to determine if their epitopes were linear (also called continuous or sequential epitopes) or assembled (also called discontinuous or conformational epitopes) 24 . As all antibodies recognized human Bak after SDS-PAGE (Fig.…”

Section: Most Bak Antibodies Have Linear Epitopesmentioning

confidence: 99%

Dissociation of Bak α1 helix from the core and latch domains is required for apoptosis

et al. 2015

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During apoptosis, Bak permeabilizes mitochondria after undergoing major conformational changes, including poorly defined N-terminal changes. Here, we characterize those changes using 11 antibodies that were epitope mapped using peptide arrays and mutagenesis. After Bak activation by Bid, epitopes throughout the a1 helix are exposed indicating complete dissociation of a1 from a2 in the core and from a6-a8 in the latch. Moreover, disulfide tethering of a1 to a2 or a6 blocks cytochrome c release, suggesting that a1 dissociation is required for further conformational changes during apoptosis. Assaying epitope exposure when a1 is tethered shows that Bid triggers a2 movement, followed by a1 dissociation. However, a2 reaches its final position only after a1 dissociates from the latch. Thus, a1 dissociation is a key step in unfolding Bak into three major components, the N terminus, the core (a2-a5) and the latch (a6-a8).

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“…Multiple synthetic N-terminally acetylated peptides were synthesized on pins (Geysen et al, 1987) using an epitope scanning kit (Cambridge Research Biochemicals) with a dilution aid (Epiguide from Labsystems) and the modifications described (McGuinness et al, 1990).…”

Section: Peptide Synthesismentioning

confidence: 99%

Two‐dimensional structure of the Opc invasin from Neisseria meningitidis

Merker

Tommassen

Kušećek

et al. 1997

Molecular Microbiology

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SummaryA two-dimensional structural model was devised for the Opc outer membrane protein invasin which contains 10 transmembrane strands and five surfaceexposed loops. One continuous epitope recognized by three monoclonal antibodies was localized to the tip of loop 2 by synthetic peptides and site-directed mutagenesis while a second, discontinuous epitope recognized by a fourth antibody was localized to loops 4 and 5 by insertion mutagenesis. These monoclonal antibodies are bactericidal and inhibit adhesion and invasion. Most of the T-cell epitopes defined by Wiertz et al. (1996) were localized to the transmembrane strands. Oligonucleotides encoding a foreign epitope (ٌ) from Semliki Forest virus were inserted into Bgl II restriction sites created by site-directed mutagenesis. The ٌ epitopes inserted in all five predicted loops were recognized on the cell surface of live Escherichia coli bacteria by a monoclonal antibody and are exposed while ٌ epitopes in the N-terminus or three predicted turns were not. The results thus confirm important predictions of the model and define five permissive sites within surface-exposed loops which can be used to insert foreign epitopes.

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Strategies for epitope analysis using peptide synthesis

Cited by 853 publications

References 25 publications

Identification of RB18A, a 205 kDa new p53 regulatory protein which shares antigenic and functional properties with p53

Identification of RB18A, a 205 kDa new p53 regulatory protein which shares antigenic and functional properties with p53

Dissociation of Bak α1 helix from the core and latch domains is required for apoptosis

Two‐dimensional structure of the Opc invasin from Neisseria meningitidis

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