The Effect of Water Activity on the Association Constant and the Enthalpy of Reaction Between Lysozyme and the Specific Antibodies D1.3 and D44.1

Goldbaum, F.A.; Schwarz, Frederick P.; Eisenstein, Edward; Cauerhff, Ana; Mariuzza, Roy A.; Poljak, Roberto J.

doi:10.1002/(sici)1099-1352(199601)9:1<6::aid-jmr240>3.3.co;2-m

Cited by 21 publications

(27 citation statements)

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“…Clearly the loss of water at the interface can improve binding, and in our system, the loss of water favors increasing enthalpy and not entropy of association in the final complex. In addition, osmotic pressure experiments showed that changes in water molarity could either increase or decrease binding enthalpy of anti-lysozyme antibodies, which correlated with whether the binding of the respective antibodies was associated with exclusion or maintenance of water molecules bound to the antibody (Braden et al, 1995; Goldbaum et al, 1996). It would be interesting to compare a similar series of antibodies from germline to late-secondary to see if the germline sequence favors entropy over enthalpy when forming the complex.…”

Section: Resultsmentioning

confidence: 99%

Light chain somatic mutations change thermodynamics of binding and water coordination in the HyHEL-10 family of antibodies

Acchione

Lipschultz

DeSantis

et al. 2009

Molecular Immunology

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Thermodynamic and structural studies addressed the increased affinity due to L-chain somatic mutations in the HyHEL-10 family of affinity matured IgG antibodies, using ITC, SPR with van’t Hoff analysis, and x-ray crystallography. When compared to the parental antibody H26L26, the H26L10 and H26L8 chimeras binding to lysozyme showed an increase in favorable ΔGo of −1. 2 ± 0. 1 Kcal mol−1 and −1. 3 ± 0. 1 Kcal mol−1, respectively. Increase in affinity of the H26L10 chimera was due to a net increase in favorable enthalpy change with little difference in change in entropy compared to H26L26. The H26L8 chimera exhibited the greatest increase in favorable enthalpy but also showed an increase in unfavorable entropy change, with the result being that the affinities of both chimeras were essentially equivalent. Site-directed L-chain mutants identified the shared somatic mutation S30G as the dominant contributor to increasing affinity to lysozyme. This mutation was not influenced by H-chain somatic mutations. Residue 30L is at the periphery of the binding interface and S30G effects an increase in hydrophobicity and decrease in H-bonding ability and size, but does not make any new energetically important antigen contacts. A new 1. 2-Å structure of the H10L10-HEL complex showed changes in the pattern of both inter- and intra-molecular water bridging with no other significant structural alterations near the binding interface compared to the H26L26-HEL complex. These results highlight the necessity for investigating both the structure and the thermodynamics associated with introduced mutations, in order to better assess and understand their impact on binding. Furthermore, it provides an important example of how backbone flexibility and water-bridging may favorably influence the thermodynamics of an antibody-antigen interaction.

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Section: Resultsmentioning

confidence: 99%

Light chain somatic mutations change thermodynamics of binding and water coordination in the HyHEL-10 family of antibodies

Acchione

Lipschultz

DeSantis

et al. 2009

Molecular Immunology

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“…To gain understanding in the molecular origins of opposite solvent effects on protein-protein interactions, we focus on a pertinent example of opposite effects of glycerol on the association constants of two different antibodies with lysozyme [21]. We use surface plasmon resonance to characterize the opposite effects of glycerol on the association constants of antibody fragments D1.3 and D44.1 over a wide concentration range (0–9 molal glycerol).…”

Section: Resultsmentioning

confidence: 99%

“…The data point marked with an asterisk is derived from Goldbaum et al [21] and all other data points are determined by surface plasmon resonance.…”

Section: Resultsmentioning

confidence: 99%

“…Exponential responses of equilibrium constants K A with respect to cosolvent molality have been observed for many biomolecular reactions [21], [43]–[54], and it has been suggested that the underlying mechanisms are closely related [17]. Considering the direct relationship between solvent-protein interactions and solvent effects on protein reactions (Eq.…”

Section: Discussionmentioning

confidence: 99%

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Quantifying the Molecular Origins of Opposite Solvent Effects on Protein-Protein Interactions

2013

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Although the nature of solvent-protein interactions is generally weak and non-specific, addition of cosolvents such as denaturants and osmolytes strengthens protein-protein interactions for some proteins, whereas it weakens protein-protein interactions for others. This is exemplified by the puzzling observation that addition of glycerol oppositely affects the association constants of two antibodies, D1.3 and D44.1, with lysozyme. To resolve this conundrum, we develop a methodology based on the thermodynamic principles of preferential interaction theory and the quantitative characterization of local protein solvation from molecular dynamics simulations. We find that changes of preferential solvent interactions at the protein-protein interface quantitatively account for the opposite effects of glycerol on the antibody-antigen association constants. Detailed characterization of local protein solvation in the free and associated protein states reveals how opposite solvent effects on protein-protein interactions depend on the extent of dewetting of the protein-protein contact region and on structural changes that alter cooperative solvent-protein interactions at the periphery of the protein-protein interface. These results demonstrate the direct relationship between macroscopic solvent effects on protein-protein interactions and atom-scale solvent-protein interactions, and establish a general methodology for predicting and understanding solvent effects on protein-protein interactions in diverse biological environments.

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“…In contrast, the use of the van't Hoff equation implies that the reaction is of a simple bimolecular type, which may not be valid in this case. Furthermore, the presence of bound water at the interface between HEL and Fab D1.3 (Bhat et al, 1994;Goldbaum et al, 1996) may also have influenced the thermodynamic parameters, as was found in the case of protein-DNA complexes (Morton and Ladbury, 1996). Others studies with systems having smaller equilibrium constants than the D1.3 antibody, which would allow DG to be determined by both calorimetry and BIAcore over a wide range of temperatures, are needed to unravel the source of these discrepancies in observed DH values.…”

Section: Thermodynamic Analysis Of Binding Parametersmentioning

confidence: 99%

Measurement of antigen–antibody interactions with biosensors

et al. 1998

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The introduction in 1990 of a new biosensor technology based on surface plasmon resonance has revolutionized the measurement of antigen-antibody binding interactions. In this technique, one of the interacting partners is immobilized on a sensor chip and the binding of the other is followed by the increase in refractive index caused by the mass of bound species. The following immunochemical applications of this new technology will be described: (1) functional mapping of epitopes and paratopes by mutagenesis; (2) analysis of the thermodynamic parameters of the interaction; (3) measurement of the concentration of biologically active molecules; (4) selection of diagnostic probes.

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The Effect of Water Activity on the Association Constant and the Enthalpy of Reaction Between Lysozyme and the Specific Antibodies D1.3 and D44.1

Cited by 21 publications

References 0 publications

Light chain somatic mutations change thermodynamics of binding and water coordination in the HyHEL-10 family of antibodies

Light chain somatic mutations change thermodynamics of binding and water coordination in the HyHEL-10 family of antibodies

Quantifying the Molecular Origins of Opposite Solvent Effects on Protein-Protein Interactions

Measurement of antigen–antibody interactions with biosensors

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