ArticleProtein science : a publication of the Protein Society1999
Energetic analysis of an antigen/antibody interface: alanine scanning mutagenesis and double mutant cycles on the HyHEL-10/lysozyme interaction.
Article in Protein science : a publication of the Protein Society, 1999. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 39 papers.
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39 citing papers in PubMed.
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- Deep mutational scanning of rabies glycoprotein defines mutational constraint and antibody-escape mutations.Cell host & microbe · 2025Article
- Epitope Mapping Using Yeast Display and Next Generation Sequencing.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Towards an optimal monoclonal antibody with higher binding affinity to the receptor-binding domain of SARS-CoV-2 spike proteins from different variants.Colloids and surfaces. B, Biointerfaces · 2023Article
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- Structural and thermodynamic basis for the recognition of the substrate-binding cleft on hen egg lysozyme by a single-domain antibody.Scientific reports · 2019Article
- Back-to-Germline (B2G) Procedure for Antibody Devolution.Antibodies (Basel, Switzerland) · 2019Article
- Structural characterization of the D290V mutation site in hnRNPA2 low-complexity-domain polymers.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Elucidation of potential sites for antibody engineering by fluctuation editing.Scientific reports · 2017Article
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- Complete mapping of viral escape from neutralizing antibodies.PLoS pathogens · 2017Article
- Engineering Specificity from Broad to Narrow: Design of a β-Lactamase Inhibitory Protein (BLIP) Variant That Exclusively Binds and Detects KPC β-Lactamase.ACS infectious diseases · 2016Article
- Structural insights into Cydia pomonella pheromone binding protein 2 mediated prediction of potentially active semiochemicals.Scientific reports · 2016Article
- Hotspots in an obligate homodimeric anticancer target. Structural and functional effects of interfacial mutations in human thymidylate synthase.Journal of medicinal chemistry · 2015Article
- Understanding the molecular basis of EGFR kinase domain/MIG-6 peptide recognition complex using computational analyses.BMC bioinformatics · 2015Article
- A frustrated binding interface for intrinsically disordered proteins.The Journal of biological chemistry · 2014Article
- Applying physics-based scoring to calculate free energies of binding for single amino acid mutations in protein-protein complexes.PloS one · 2013Article
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Abstract
Alanine scanning mutagenesis of the HyHEL-10 paratope of the HyHEL-10/HEWL complex demonstrates that the energetically important side chains (hot spots) of both partners are in contact. A plot of deltadeltaG(HyHEL-10_mutant) vs. deltadeltaG(HEWL_mutant) for the five of six interacting side-chain hydrogen bonds is linear (Slope = 1). Only 3 of the 13 residues in the HEWL epitope contribute >4 kcal/mol to the free energy of formation of the complex when replaced by alanine, but 6 of the 12 HyHEL-10 paratope amino acids do. Double mutant cycle analysis of the single crystallographically identified salt bridge, D32H/K97, shows that there is a significant energetic penalty when either partner is replaced with a neutral side-chain amino acid, but the D32(H)N/K97M complex is as stable as the WT. The role of the disproportionately high number of Tyr residues in the CDR was evaluated by comparing the deltadeltaG values of the Tyr --> Phe vs. the corresponding Tyr --> Ala mutations. The nonpolar contacts in the light chain contribute only about one-half of the total deltadeltaG observed for the Tyr --> Ala mutation, while they are significantly more important in the heavy chain. Replacement of the N31L/K96 hydrogen bond with a salt bridge, N31D(L)/K96, destabilizes the complex by 1.4 kcal/mol. The free energy of interaction, deltadeltaG(int), obtained from double mutant cycle analysis showed that deltadeltaG(int) for any complex for which the HEWL residue probed is a major immunodeterminant is very close to the loss of free energy observed for the HyHEL-10 single mutant. Error propagation analysis of double mutant cycles shows that data of atypically high precision are required to use this method meaningfully, except where large deltadeltaG values are analyzed.
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