Hydrogen exchange analysis of ligand-induced conformational changes in Fab

Liberti, Paul A.; Bausch, Dulcie M.; Chu, M.S.Y.

doi:10.1021/bi00507a056

Cited by 7 publications

(2 citation statements)

References 20 publications

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“…In some studies (16,17), no conformational changes were observed upon binding of a small ligand. In others, conformational alterations correlated with hapten or antigen binding have been inferred on the basis of increased conformational stability ofthe Fab (18,19), decreased solubility (20), changes in the accessibility of bound hydrogens for solvent exchange (21), and results of numerous spectroscopic studies (22)(23)(24)(25)(26). In crystallographically based studies of conformational changes in the light-chain dimer Mcg, it was concluded that ligand binding caused alterations in both VL-VL and CL-CL interactions (27,28).…”

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confidence: 99%

Dual conformations of an immunoglobulin light-chain dimer: heterogeneity of antigen specificity and idiotope profile may result from multiple variable-domain interaction mechanisms.

Stevens

Chang

Schiffer

1988

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

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The structure of an immunoglobulin antigenbinding fragment (Fab) has been thought to be invariantly defined by well-coiserved amino acid residues in the variable domains of the heavy and light chains. These conserved residues enable folding of the polypeptide segments into the characteristic immunoglobulin fold domains and are the major controllers of interactions between domains. However, crystallographic studies of some immunoglobulin light-chain dimers have suggested and the crystallographic structure of the Fab in an Fab-neuraminidase complex may have proven that antibodies are not restricted to a single, invariant relative positioning of the two variable domains. We propose that in some cases the detailed quaternary structural relationships between the variable domains of heavy and light chains are not restricted to those of the canonical Fab. It is unclear whether alterations of these relationships occur only after complex formation with antigen or, if in ligand-free solution, Fab conformers might coexist in relative concentrations determined by isomerization rates. In the latter case, antibody-presenting lymphocytes may be polyspecific, and the specificity of lymphocytes might be modulated by anti-idiotopic antibodies complexed to cell surface receptors. In either case, the idiotopic repertoire displayed by an antibody or lymphocyte surface receptor might be changed by the presence or absence of antigen.Crystallographic studies of antigen-binding fragments (Fabs) have indicated that the principal determinants of quaternary structure are highly conserved domain-domain interactions (1)(2)(3)(4)(5) and that functional diversity originates from residue alternations, insertions, and deletions in segments of the heavy (H)-and light (L)-chain variable (V) domains designated hypervariable or complementarity determining regions (6). Several analogous studies with light-chain dimers have also supported the monomorphic conformation concept (7-10). However, in two instances (11,12), the observed Bence Jones protein conformations were anomalous; V-V domain interactions were found that had not been observed in Fabs. Although the relationships between these V domains were of significant interest in terms of the physical chemistry of antibody domain interactions, the physiological implications of the structures of the light-chain dimers formed by proteins Rhe (11)

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mentioning

confidence: 99%

Dual conformations of an immunoglobulin light-chain dimer: heterogeneity of antigen specificity and idiotope profile may result from multiple variable-domain interaction mechanisms.

Stevens

Chang

Schiffer

1988

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

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“…Like HDX, the sister technique of HTX, pioneered by Englander and coworkers (Englander, 1963), was among the first to be deployed for antigen-antibody interface analysis (Ashman et al, 1971;Liberti et al, 1972;Liberti et al, 1981). As other detection methods evolved, HDX was coupled with infrared spectroscopy (Závodszky et al, 1981), subsequently with NMR for epitope mapping (Paterson et al, 1990).…”

Section: Hdx For Epitope Mapping 41 Hdx Theory and Usementioning

confidence: 99%

Hydrogen deuterium exchange and other mass spectrometry- based approaches for epitope mapping

Jethva

Gross

2023

Front. Anal. Sci.

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Antigen-antibody interactions are a fundamental subset of protein-protein interactions responsible for the “survival of the fittest.” Determining the interacting interface of the antigen, called an epitope, and that on the antibody, called a paratope, is crucial to antibody development. Because each antigen presents multiple epitopes (unique footprints), sophisticated approaches are required to determine the target region for a given antibody. Although X-ray crystallography, Cryo-EM, and nuclear magnetic resonance can provide atomic details of an epitope, they are often laborious, poor in throughput, and insensitive. Mass spectrometry-based approaches offer rapid turnaround, intermediate structural resolution, and virtually no size limit for the antigen, making them a vital approach for epitope mapping. In this review, we describe in detail the principles of hydrogen deuterium exchange mass spectrometry in application to epitope mapping. We also show that a combination of MS-based approaches can assist or complement epitope mapping and push the limit of structural resolution to the residue level. We describe in detail the MS methods used in epitope mapping, provide our perspective about the approaches, and focus on elucidating the role that HDX-MS is playing now and in the future by organizing a discussion centered around several improvements in prototype instrument/applications used for epitope mapping. At the end, we provide a tabular summary of the current literature on HDX-MS-based epitope mapping.

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Effect of reduction and alkylation on structure and function of rabbit IgG antibody—II. Effects on classical pathway C3 convertase formation

Johnson

Hoffmann

1984

Molecular Immunology

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Hydrogen exchange analysis of ligand-induced conformational changes in Fab

Cited by 7 publications

References 20 publications

Dual conformations of an immunoglobulin light-chain dimer: heterogeneity of antigen specificity and idiotope profile may result from multiple variable-domain interaction mechanisms.

Dual conformations of an immunoglobulin light-chain dimer: heterogeneity of antigen specificity and idiotope profile may result from multiple variable-domain interaction mechanisms.

Hydrogen deuterium exchange and other mass spectrometry- based approaches for epitope mapping

Effect of reduction and alkylation on structure and function of rabbit IgG antibody—II. Effects on classical pathway C3 convertase formation

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