Spectroscopic, Calorimetric, and Kinetic Demonstration of Conformational Adaptation in Peptide-Antibody Recognition

Leder, Lukas; Berger, Christopher L.; Bornhauser, Susanne; Wendt, H.; Ackermann, Friederike; Jelesarov, Ilian; Bosshard, Hans Rudolf

doi:10.1021/bi00050a035

Cited by 71 publications

(71 citation statements)

References 27 publications

(50 reference statements)

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“…This value is not far from To saturate the column-bound antibody, peptide LZ had to be circulated through the column for several hours, whereas 30 min sufficed for saturation with the cognate peptide LZ7P14P. This confirmed the slower reaction of 42PF with the non-cognate antigen observed previously [4]. After elution of the excess unbound antigen, 7-8 nmol peptide/ml affinity gel were adsorbed and could not be eluted by washing with several column volumes of The antigen .…”

Section: Resultssupporting

confidence: 78%

“…The cognate peptide LZ7P14P (against which antibody 42PF was raised) binds with a AG value of -43.5 ? 1 .S kJ/mol [4]. Hence, antibody-promoted unfolding of the triple-stranded coiled coil seems feasible, even considering that peptide LZ binds less strongly than the cognate peptide LZ7P14P.…”

Section: Discussionmentioning

confidence: 99%

“…Substitution of Pro7 by Lys and Pro14 by Ala produces peptide LZ, and this peptide associates to an all-parallel, triple-stranded coiled coil [8, 91. The antigenic cross-reactivity of antibody 42PF with peptide LZ was considerable in view of the different conformations of peptides LZ7P14P (the cognate antigen) and LZ (the non-cognate antigen). In a conformation-sensitive competition immunoassay [lo], 20 pM LZ reduced by 50% the binding of 0.3 pM LZ7P14P to 0.3 pM antibody 42PF [4]. Furthermore, the fluorescence of a 7-dimethylaminocoumarin group attached to the N terminus of LZ was strongly quenched in the triple-stranded coiled-coil conformation of peptide LZ but increased when the peptide was bound to antibody 42PF [4, 91. In addition, the reaction of 42PF with non-cognate LZ was much slower than with cognate LZ7P14P.…”

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confidence: 99%

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A Monoclonal Antibody Induces Opening of a Coiled Coil

Dürr

Bosshard

1997

European Journal of Biochemistry

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Antibody specificity is limited, and different antigens may cross‐react to varying degrees with the same antibody. An example is monoclonal antibody 42PF elicited against a 29‐residue random‐coil pep‐tide. 42PF cross‐reacts with a triple‐stranded coiled coil of related amino acid sequence, and there was circumstantial evidence for an antibody‐induced opening of the coiled coil [Leder, L., Berger, C., Bornhauser, S., Wendt, H., Ackermann, F., Jelesarov, I. & Bosshard, H. R. (1995) Biochemistry 34, 16509–16518]. To reveal in a direct way that antibody 42PF induces the opening of the coiled coil, we have compared the rates of H/D exchange of amide protons in the free and in the antibody‐bound coiled‐coil peptide using electrospray‐ionization mass spectrometry for the analysis of deuterium incorporation. Complete H/D exchange lasted several days in the tightly folded coiled‐coil conformation while in the presence of antibody 42PF the overall exchange took only minutes, corresponding to a thousand‐fold decrease of protection of the most slowly exchanging amide hydrogens of the folded coiled coil. This demonstrates that the antibody induced a considerable opening of the coiled coil.

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Section: Resultssupporting

confidence: 78%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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A Monoclonal Antibody Induces Opening of a Coiled Coil

Dürr

Bosshard

1997

European Journal of Biochemistry

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“…1): (i) the induced fit model and (ii) the conformational selection/population shift model (7)(8)(9)(10)(11)(12)(13)(14). As defined originally by Koshland (1), the induced fit model states that the achievement of the final key-lock state, that is, the precise orientation of catalytic groups and residues necessary for catalysis, occurs only after changes in protein structure that are induced by the binding of a substrate.…”

mentioning

confidence: 99%

Enzymes with lid-gated active sites must operate by an induced fit mechanism instead of conformational selection

Sullivan

Holyoak

2008

Proc. Natl. Acad. Sci. U.S.A.

147

217

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The induced fit and conformational selection/population shift models are two extreme cases of a continuum aimed at understanding the mechanism by which the final key-lock or active enzyme conformation is achieved upon formation of the correctly ligated enzyme. Structures of complexes representing the Michaelis and enolate intermediate complexes of the reaction catalyzed by phosphoenolpyruvate carboxykinase provide direct structural evidence for the encounter complex that is intrinsic to the induced fit model and not required by the conformational selection model. In addition, the structural data demonstrate that the conformational selection model is not sufficient to explain the correlation between dynamics and catalysis in phosphoenolpyruvate carboxykinase and other enzymes in which the transition between the uninduced and the induced conformations occludes the active site from the solvent. The structural data are consistent with a model in that the energy input from substrate association results in changes in the free energy landscape for the protein, allowing for structural transitions along an induced fit pathway.enzyme dynamics ͉ population shift ͉ phosphoenolpyruvate carboxykinase I t has been 50 years since the original presentation of the induced fit hypothesis by Daniel Koshland (1) that built upon Emil Fisher's key-lock principle (2) and introduced the idea that the inherent dynamic properties of enzymes play an essential role in the processes of molecular recognition and catalysis. Over the last decade, with the advent of new instrumentation and novel experimental approaches there has been a resurgence in investigations focused upon understanding the specific ways in that these dynamic properties influence the ability of enzymes to achieve enormous levels of selectivity and catalytic power (refs. 3-6 and references therein).Presently there are two primary models used to explain the mechanism by which an enzyme can move toward adopting the final key-lock state ( Fig. 1): (i) the induced fit model and (ii) the conformational selection/population shift model (7-14). As defined originally by Koshland (1), the induced fit model states that the achievement of the final key-lock state, that is, the precise orientation of catalytic groups and residues necessary for catalysis, occurs only after changes in protein structure that are induced by the binding of a substrate. This pathway is represented by the equilibrium constants K 1 and K 2 in Fig. 1. Intrinsic to this model is the ability of the enzyme to form an ''encounter complex'' † (Fig. 1B). In this complex the enzyme is liganded identically to the eventual catalytically competent key-lock state; however, the conformational change leading to the active key-lock state has not yet occurred. In contrast, the conformational selection model states that the enzyme exists in multiple conformational states, of which the substrate has a high affinity for the active or key-lock state. In this model the ligand is suggested to bind to this lowly populated high-energy ...

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“…In a leucine zipper model, the antibody selected a pre-existing monomeric conformer and shifted the equilibrium from a native coiled-coil to a monomeric, unfolded species (11,12). A somehow different example comes from an epitope within the folded dimeric transcriptional regulator E2 from human papillomavirus (HPV).…”

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confidence: 99%

Minute Time Scale Prolyl Isomerization Governs Antibody Recognition of an Intrinsically Disordered Immunodominant Epitope

Fassolari

Chemes

Gallo

et al. 2013

Journal of Biological Chemistry

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Background: Kinetic discrimination is essential in antibody⅐antigen recognition. Results: An otherwise fast second-order reaction is limited by a slow proline isomerization of the epitope. Conclusion: The antibody recognizes the less populated isoform, suggesting presentation of the epitope as a non-native conformer. Significance: The conformational diversity of an intrinsically disordered viral epitope slows down the reaction with the antibody without specificity cost.

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Spectroscopic, Calorimetric, and Kinetic Demonstration of Conformational Adaptation in Peptide-Antibody Recognition

Cited by 71 publications

References 27 publications

A Monoclonal Antibody Induces Opening of a Coiled Coil

A Monoclonal Antibody Induces Opening of a Coiled Coil

Enzymes with lid-gated active sites must operate by an induced fit mechanism instead of conformational selection

Minute Time Scale Prolyl Isomerization Governs Antibody Recognition of an Intrinsically Disordered Immunodominant Epitope

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