Structural and physicochemical analysis of the reaction between the anti‐lysozyme antibody D1.3 and the anti‐idiotopic antibodies E225 and E5.2

Tello, Diana; Eisenstein, Edward; Schwarz, Frederick P.; Goldbaum, Fernando A.; Fields, Barry A.; Mariuzza, Roy A.; Poljak, Roberto J.

doi:10.1002/jmr.300070108

Cited by 22 publications

(9 citation statements)

References 24 publications

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“…So what causes the reduction in the k on ? Previously, kinetic and structure studies of the anti-lysozyme antibody D1.3 have indicated that local conformational adaptation is responsible for the association rate (40). Furthermore, statistical thermodynamic analysis of the same antibody has shown that the binding effects can propagate to a subset of residues at distal sites when the binding sites include the region with conformational fluctuations (41).…”

Section: Discussionmentioning

confidence: 99%

Kinetic Interaction Analysis of Human Interleukin 5 Receptor α Mutants Reveals a Unique Binding Topology and Charge Distribution for Cytokine Recognition

Ishino

Pasut

Scibek

et al. 2004

Journal of Biological Chemistry

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Human interleukin 5 receptor ␣ (IL5R␣) comprises three fibronectin type III domains (D1, D2, and D3) in the extracellular region. Previous results have indicated that residues in the D1D2 domains are crucial for high affinity interaction with human interleukin 5 (IL5). Yet, it is the D2D3 domains that have sequence homology with the classic cytokine recognition motif that is generally assumed to be the minimum cytokinerecognizing unit. In the present study, we used kinetic interaction analysis of alanine-scanning mutational variants of IL5R␣ to define the residues involved in IL5 recognition. Soluble forms of IL5R␣ variants were expressed in S2 cells, selectively captured via their C-terminal V5 tag by anti-V5 tag antibody immobilized onto the sensor chip and examined for IL5 interaction by using a sandwich surface plasmon resonance biosensor method. Marked effects on the interaction kinetics were observed not only in D1 (Asp 55 , Asp 56 , and Glu 58 ) and D2 (Lys 186 and Arg 188 ) domains, but also in the D3 (Arg 297 ) domain. Modeling of the tertiary structure of IL5R␣ indicated that these binding residues fell into two clusters. The first cluster consists of D1 domain residues that form a negatively charged patch, whereas the second cluster consists of residues that form a positively charged patch at the interface of D2 and D3 domains. These results suggest that the IL5⅐IL5R␣ system adopts a unique binding topology, in which the cytokine is recognized by a D2D3 tandem domain combined with a D1 domain, to form an extended cytokine recognition interface. Interleukin 5 (IL5)1 is a T cell-derived cytokine that plays a central role in maturation and proliferation of eosinophils (1). IL5 exerts biological functions through recruitment of a cell surface receptor composed of two polypeptide chains (2), ␣ and ␤. The ␣ chain is IL5-specific and is called IL5 receptor ␣ (IL5R␣), whereas the ␤ chain is shared with IL3 and GM-CSF (3, 4) and is called common ␤ chain (␤c). The ␣ chains for IL3, GM-CSF, and IL5R␣ also share a high degree of amino acid sequence similarity and constitute a distinct subgroup within the cytokine receptor family (5). Because IL5 has been implicated in the pathology of eosinophil-related inflammatory diseases, designing specific antagonists for IL5R␣ may offer therapeutic benefits in the treatment of such diseases (6, 7).IL5 initially binds to IL5R␣ with high affinity, and the resulting complex recruits ␤c to induce cytoplasmic signal transduction. Human IL5R␣ alone binds IL5 with an equilibrium dissociation constant (K d ) of 0.3-0.6 nM when expressed in COS cells (8). The binding affinity is increased only 2-to 5-fold when human ␣ and ␤ chains are co-expressed (3). In other words, IL5R␣ provides most of the IL5 binding energy, whereas ␤c is essentially for signaling. IL5R␣ consists of an extracellular region, a single transmembrane region and a cytoplasmic region. A soluble form of IL5R␣ (sIL5R␣) containing only the extracellular region was cloned and expressed (9). This form has provided a conv...

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Section: Discussionmentioning

confidence: 99%

Kinetic Interaction Analysis of Human Interleukin 5 Receptor α Mutants Reveals a Unique Binding Topology and Charge Distribution for Cytokine Recognition

Ishino

Pasut

Scibek

et al. 2004

Journal of Biological Chemistry

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“…As for affinity studies, a number of data are now available on kinetic measurements of antigen-antibody interactions assessed by surface plasmon resonance (8,(19)(20)(21)(22). These studies show that equilibrium constants of these interactions vary from 10 6 to 10 11 with k ass values ranging from 10 3 to 10 6 and k diss ranging from 10 Ϫ3 to 10…”

Section: Discussionmentioning

confidence: 99%

Can immunoglobulin C(H)1 constant region domain modulate antigen binding affinity of antibodies?

Pritsch¹,

Hudry-Clergeon²,

Buckle³

et al. 1996

J. Clin. Invest.

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Although the switch process is frequently associated with affinity maturation, the constant region is not assumed to play a role in Ag-Ab binding. In the present work, we demonstrate that two clonally related human monoclonal Igs sharing identical V H and V L sequences, but expressing different isotypes (IgA1 PER and IgG1 PER ), bind tubulin with significantly different affinities. This difference was mainly accounted for by a disparity in the association rate constants. These results suggest that affinity maturation of this clone could be achieved through class switching in the absence of further somatic mutations. Since the differences observed were found at the Fab level, they also suggest a role for the C H 1 domain in structuring the Ag-binding site into a more kinetically competent form. ( J. Clin. Invest. 1996. 98:2235-2243.)

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“…An alternative possible explanation for the effect of mutations on the association state might be differential solvation or desolvation. It is known that solvent structure can exert a significant effect in macromolecular interactions (45). Solvent structure associated with the unusually exposed phenyl group of Phe-43 could be quite different from that with its valyl counterpart, and its rate of reorganization on binding to gp120 could then also be different.…”

Section: Discussionmentioning

confidence: 99%

Kinetic and structural analysis of mutant CD4 receptors that are defective in HIV gp120 binding

Myszka

Tendian

et al. 1996

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

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The T-cell antigen coreceptor CD4 also serves as the receptor for the envelope glycoprotein gp120 of HIV. Extensive mutational analysis of CD4 has implicated residues from a portion of the extracellular amino-terminal domain (D1) in gp120 binding. However, none of these proteins has been fully characterized biophysically, and thus the precise effects on molecular structure and binding interactions are unknown. In the present study, we produced soluble versions of three mutant CD4 molecules (F43V, G47S, and A55F) and characterized their structural properties, thermostability, and ability to bind gp120. Crystallographic and thermodynamic analysis showed minimal structural alterations in the F43V and G47S mutant proteins, which have solvent-exposed mutant side chains. In contrast, some degree of disorder appears to exist in the folded state of A55F, as a result of mutating a buried side chain. Real time kinetic measurements of the interaction of the mutant proteins with gp120 showed affinity decreases of 5-fold for G47S, 50-fold for A55F, and 200-fold for F43V. Although both rate constants for the binding reaction were affected by these mutations, the loss in affinity was mainly due to a decrease in on rates, with less drastic changes occurring in the off rates. These observations suggest the involvement of conformational adaptation in the CD4-gp120 interaction. Together, the structural and kinetic data confirm that F43V is a critical residue in gp120 recognition site, which may also include main chain interactions at residue Gly-47.

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Structural and physicochemical analysis of the reaction between the anti‐lysozyme antibody D1.3 and the anti‐idiotopic antibodies E225 and E5.2

Cited by 22 publications

References 24 publications

Kinetic Interaction Analysis of Human Interleukin 5 Receptor α Mutants Reveals a Unique Binding Topology and Charge Distribution for Cytokine Recognition

Kinetic Interaction Analysis of Human Interleukin 5 Receptor α Mutants Reveals a Unique Binding Topology and Charge Distribution for Cytokine Recognition

Can immunoglobulin C(H)1 constant region domain modulate antigen binding affinity of antibodies?

Kinetic and structural analysis of mutant CD4 receptors that are defective in HIV gp120 binding

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