Folding and immunogenicity of a loop-structured peptide using the zinc-finger motif

Kobs-Conrad, Susan F.; DiGeorge, A. M.; Lee, Hyosil; Kaumaya, Pravin T. P.

doi:10.1007/978-94-011-1470-7_252

Cited by 10 publications

(12 citation statements)

References 2 publications

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“…In the past, attempts to engineer antigenic determinants have focussed mainly on super secondary structures [12][13][14][15]. While these strategies have their own merits, one drawback lies in the fact that the designed sequences are quite long (often in the range of 35-80 residues).…”

Section: Resultsmentioning

confidence: 99%

“…Transferred NOE [6][7][8][9] and crystallographic [10,11] studies of peptides complexed to Fab fragments of monoclonal antibodies have clearly demonstrated the formation of ordered backbone conformations in the bound states. The incorporation of specific stereochemical constraints into an antigenic peptide that stabilize conformational states recognized by antibodies, appears to provide an attractive approach to engineering improved immunological characteristics [12][13][14][15]. We describe in this report that helix stabilization in a 21 residue synthetic peptide fragment of chicken riboflavin carrier protein (cRCP) dramatically enhances the immunogenic properties of the peptide.…”

Section: Introductionmentioning

confidence: 99%

“…The observation that antibodies raised to short synthetic peptide fragments often cross react with the native protein antigens has stimulated a great deal of interest in the development of peptide vaccines [14]. Well-defined conformational states have been demonstrated for a wide variety of antigenic peptides in solution [5].…”

Section: Introductionmentioning

confidence: 99%

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Enhancing peptide antigenicity by helix stabilization

et al. 1995

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The engineering of antigenic determinants on super secondary structures using de novo design approaches often involves synthesis of long peptide chains (35--80 residues long). This communication illustrates that the stabilization of secondary structure by rational design can also greatly enhance immunogenicity and antigenicity, but in much shorter peptide sequences (21 residues long). A peptide epitope the sequence of which has been derived from the C-terminus of the chicken riboflavin carrier protein (cRCP), H2N-Tyr-His-Ala-Cys-Gln-Lys-Lys-Leu-LeuL~Phe-GIu-AIa-Lm-GIn-GIn-GIu-Giu-GIy-GIu-GIu-OH, has been chosen for analysis. Helical conformations were induced in the peptide in aqueous trifluoroetbanol. Analogs were designed to stabilize this conformation in water by either the introduction of appropriately spaced ion pairs or the strongly helix nucleating residue a-aminoisobutyric acid (Aib), substituted for Ala/Gly, thus affording a comparison of the helix stabilization strategies. Circular dichroism (CD) results demonstrate that all the designed analogs are appreciably more helical than the parent peptide in 50% aqueous trifluoroethanol. Peptide antisera were raised for all analogs in rabbits. The affinities of these antisera for the native protein antigen, determined using a chaotrope disrupted binding assay, correlated very well with the helix content determined by CD.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Enhancing peptide antigenicity by helix stabilization

et al. 1995

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“…1. We have previously demonstrated that the design, synthesis, and immunological and structural characterization of such motifs can be achieved successfully (33). The peptide of p-MVFGSC90 was synthesized as chimeric constructs incorporating an 18-residue promiscuous T-helper measles virus (MVF sequence, 288 KLLSLIKGVIVHRLEGVE 305 ) T cell epitope linked via a 4-residue linker (GPSL) and p-GSC90 B cell epitope on a Milligen/Biosearch 9600 solid-phase peptide synthesizer as described under "Experimental Procedures."…”

Section: Generation Of Antibody Against the Epitope Of Glutathionementioning

confidence: 99%

Protein Tyrosine Nitration of the Flavin Subunit Is Associated with Oxidative Modification of Mitochondrial Complex II in the Post-ischemic Myocardium

Chen

Rawale

et al. 2008

Journal of Biological Chemistry

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Increased O 2. and NO production is a key mechanism of mito- To test for protein nitration, rats were subjected to 30 min of coronary ligation followed by 24 h of reperfusion. Tissue homogenates were immunoprecipitated with AbGSC90 and probed with antibodies against 3-nitrotyrosine. Enhancement of protein tyrosine nitration was detected in the post-ischemic myocardium. Isolated SQR was subjected to in vitro protein nitration with peroxynitrite, leading to site-specific nitration at the 70-kDa polypeptide and impairment of SQR electron transfer activity. Protein nitration of SQR further impaired its protein-protein interaction with Complex III. Liquid chromatography/tandem mass spectrometry analysis indicated that Tyr-56 and Tyr-142 were involved in protein tyrosine nitration. When the isolated SQR was subjected to in vitro S-glutathionylation, oxidative modification and impairment mediated by peroxynitrite were significantly decreased, thus confirming the protective effect of S-glutathionylation from the oxidative damage of nitration.

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“…In this approach, a core ␤-sheet template consisting of alternating Gly/Lys is synthesized such that the ⑀ side chains of the lysine residues allow for the growth of individual epitopes in varying permutations. Model peptides incorporating different combinations of B-cell and T-cell epitopes from either the lactate dehydrogenase C 4 (LDH-C 4 ) antigen or the human T-cell leukemia virus type 1 (HTLV-1) 1 envelope surface-exposed subunit synthesized in a template format resulted in enhanced immunogenicity in several inbred and outbred strains of mice and rabbits eliciting antibodies reactive with the native protein (4,(7)(8)(9).…”

mentioning

confidence: 99%

De Novo Design of Peptide Immunogens That Mimic the Coiled Coil Region of Human T-cell Leukemia Virus Type-1 Glycoprotein 21 Transmembrane Subunit for Induction of Native Protein Reactive Neutralizing Antibodies

Sundaram

Lynch

Rawale

et al. 2004

Journal of Biological Chemistry

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Peptide vaccines able to induce high affinity and protective neutralizing antibodies must rely in part on the design of antigenic epitopes that mimic the threedimensional structure of the corresponding region in the native protein. We describe the design, structural characterization, immunogenicity, and neutralizing potential of antibodies elicited by conformational peptides derived from the human T-cell leukemia virus type 1 (HTLV-1) gp21 envelope glycoprotein spanning residues 347-374. We used a novel template design and a unique synthetic approach to construct two peptides (WCCR2T and CCR2T) that would each assemble into a triple helical coiled coil conformation mimicking the gp21 crystal structure. The peptide B-cell epitopes were grafted onto the ⑀ side chains of three lysyl residues on a template backbone construct consisting of the sequence acetyl-XGKGKGKGCONH2 (where X represents the tetanus toxoid promiscuous T cell epitope (TT) sequence 580 -599). Leucine substitutions were introduced at the a and d positions of the CCR2T sequence to maximize helical character and stability as shown by circular dichroism and guanidinium hydrochloride studies. Serum from an HTLV-1-infected patient was able to recognize the selected epitopes by enzyme-linked immunosorbent assay (ELISA). Mice immunized with the wild-type sequence (WCCR2T) and the mutant sequence (CCR2T) elicited high antibody titers that were capable of recognizing the native protein as shown by flow cytometry and whole virus ELISA. Sera and purified antibodies from immunized mice were able to reduce the formation of syncytia induced by the envelope glycoprotein of HTLV-1, suggesting that antibodies directed against the coiled coil region of gp21 are capable of disrupting cell-cell fusion. Our results indicate that these peptides represent potential candidates for use in a peptide vaccine against HTLV-1.The efficacy of potential peptide vaccines for bacterial, parasitic, and viral diseases as well as cancer depends partly on the induction of broadly protective, high affinity neutralizing antibody responses. Such responses rely on the design of epitopes able to mimic the three-dimensional structure of the respective B cell epitope in the native protein as well as the activation of the helper T cell component of the immune response. Over the last several years, our laboratory has conceptualized and implemented several approaches to engineer conformational epitopes in tandem with a chimeric strategy to deliver potential vaccine candidates. The design involves selecting surface-oriented antigenic epitopes that display certain secondary structural attributes (␣-helix, ␤-turn, and loop) that are then linked to a "promiscuous" T-helper cell epitope via a four-residue turn sequence (GPSL). These chimeric peptides allow independent folding of the B-cell and T-cell epitope and are very effective in overcoming major histocompatibility complex restriction in various strains of mice and in eliciting high titered and high affinity antibody responses (1-4). Additionally...

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Folding and immunogenicity of a loop-structured peptide using the zinc-finger motif

Cited by 10 publications

References 2 publications

Enhancing peptide antigenicity by helix stabilization

Enhancing peptide antigenicity by helix stabilization

Protein Tyrosine Nitration of the Flavin Subunit Is Associated with Oxidative Modification of Mitochondrial Complex II in the Post-ischemic Myocardium

De Novo Design of Peptide Immunogens That Mimic the Coiled Coil Region of Human T-cell Leukemia Virus Type-1 Glycoprotein 21 Transmembrane Subunit for Induction of Native Protein Reactive Neutralizing Antibodies

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