Conformational flexibility of an antibody combining site composed of two identical V regions

Gavish, M.; Dwek, Raymond A.

doi:10.1002/eji.1830080109

Cited by 7 publications

(7 citation statements)

References 31 publications

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“…This demonstrates that binding at only one site per VL dimer is affecting the protein resonances, in agreement with the equilibrium dialysis results, at pH 5.0, ofGavish et al (1978). If binding were occurring at two sites, many or all of the protein resonances would have titrated over less than 50% of their total range.…”

supporting

confidence: 88%

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Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Wr¹,

Rj²,

Gavish³

et al. 1981

Biochemistry

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The binding of dinitrophenyl (DNP) compounds to the VL dimer of protein 315 (IgA/X2) results in small perturbations of about ten resonances in the aromatic region of the 270-MHz 'H NMR spectrum of the protein. From a comparison of the chemical shifts of these resonances with the chemical shifts of resonances which are affected by the binding of DNP compounds to the Fv fragment of protein 315, it is concluded that the conformation of combining site residues in the VL domain of the Fv fragment is maintained in the VL dimer. The binding of DNP compounds to the VL dimer is therefore considered to reflect the retention of structural features important in determining the specificity of the Fv fragment and not the fortuitious creation of a binding site. These findings provide a structural explanation for the observation that antibodies with a X2 light chain are contained in the anti-DNP response of BALB/c mice [Cotner, T., &

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supporting

confidence: 88%

“…It is expected that the DNP ring will stack between the two T r~-9 3~ residues. Evidence for a stacking interaction with a tryptophan in the combining site of the L-chain dimer has been obtained by CD (Freed et al, 1976) and by UV difference spectroscopy (Gavish et al, 1978).…”

Section: Retention Of Combining Site Conformation In the Separatedmentioning

confidence: 99%

Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Wr¹,

Rj²,

Gavish³

et al. 1981

Biochemistry

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“…Like Mcg L,, and in contrast to Fv315, VL315 seems to be endowed with conformational flexibility. (V,), undergoes a marked transition between two conformations near pH 7 [2]. The two conformers differ significantly by the number of ligands that can be accommodated to the site; two ligands for the high pH conformer and one for the low pH conformer.…”

Section: Resultsmentioning

confidence: 99%

“…affinity of Fv are the same whether anti-V, antibodies were present or absent in the experiment. Since at pH 5.0, (VL), possess only one site for DNP-lysine [2], we have tested the effect of anti-V, Fab' on the binding of DNP-lysine by (V,), and Fv. In contrast to the effect of anti-V, Fab'onthebinding properties of VL at pH 8.0, no effect was found at pH 5.0.…”

Section: Equilibrium Dialysismentioning

confidence: 99%

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Anti‐V region framework antibodies affect the ligand binding of V_L dimer

Ben‐Neriah

Gavish

1979

Eur J Immunol

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The effect of antibodies to the light chain variable region (VL) of protein MOPC-315 (alpha, lambda 2), on the binding of hapten by VL315 dimer or Fv315 (VL + VH) was studied by equilibrium dialysis. Anti-VL did not change the binding properties of Fv but affected the binding properties of VL dimer. At pH 5, the binding properties of VL in the presence or absence of anti-VL were the same, whereas at pH 8, anti-VL reduced the number of ligands bound to VL from two to one. It has previously been shown that VL dimer binds one ligand at pH 5 and two ligands at pH 8, and that VL conformation at pH 5 is tighter. Hence, our results suggest that anti-VL tightens the conformation of VL dimer at pH 8.0 such that it can bind only one ligend. Since Fv is not affected by anti-VL, the results indicate that a combining site made of two identical chains (VL dimer) can undergo a conformational change upon interaction with its antibody. Such conformational change can indirectly affect the binding properties.

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Antigenic markers of V domain of mouse MOPC 21 IgG₁ L chain. The conformational basis of V_L domain markers and content of MOPC 21 V_L‐like immunoglobulins in sera of inbred mouse strains

et al. 1983

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Antisera were raised in rabbits against L chains, isolated from mouse myeloma protein MOPC21 (gamma 1, chi V chi 15 group). Specific antibodies for the V and C domain of MOPC21 L chain were obtained by cross-immunoadsorption of the antisera. The pure anti-V and anti-C antibodies were fixed on diazocellulose and used as immunosorbents. The inhibitory capacity of L chin-monomers and dimers isolated from the L chain preparation was compared to that of intact IgG1 using binding inhibition of 125I-labeled IgG1 on the antibody-containing immunosorbents. It was established that changes of IgG1 quaternary structure influences the conformational state of the L chain V domain only. The inhibitory capacity of the V domain is 1000-fold lower in L monomers, if compared with native IgG1, and only 10-fold lower than in L dimers. The inhibiting capacity of the C domain, however, does not differ in L monomers and intact IgG1. Thus the conformational rigidity of the C domain co-exists with conformational flexibility of the V domain on the same polypeptide chain. We tried to estimate the content of MOPC21 V1-like normal IgG in mouse serum of 6 inbred strains using antibodies against the V1 domain. Data obtained by inhibition of radioimmunoadsorption, indicate that in C57BL/6 mice 0.08% of normal serum Ig carries a V1 region which is idiotypically related to the V1 of MOPC21. In serum Ig of BALB/c mice the percentage is 0.16.

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Conformational flexibility of an antibody combining site composed of two identical V regions

Abstract: Conformational flexibility of an antibody combining site composed of two identical V regions"

Cited by 7 publications

References 31 publications

Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Anti‐V region framework antibodies affect the ligand binding of V_L dimer

Antigenic markers of V domain of mouse MOPC 21 IgG₁ L chain. The conformational basis of V_L domain markers and content of MOPC 21 V_L‐like immunoglobulins in sera of inbred mouse strains

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Conformational flexibility of an antibody combining site composed of two identical V regions

Abstract: Conformational flexibility of an antibody combining site composed of two identical V regions"

Cited by 7 publications

References 31 publications

Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Antibody specificity: a 270-MHz hydrogen-1 nuclear magnetic resonance study of the binding of dinitrophenyl compounds to the VL dimer of protein 315

Anti‐V region framework antibodies affect the ligand binding of VL dimer

Antigenic markers of V domain of mouse MOPC 21 IgG1 L chain. The conformational basis of VL domain markers and content of MOPC 21 VL‐like immunoglobulins in sera of inbred mouse strains

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Anti‐V region framework antibodies affect the ligand binding of V_L dimer

Antigenic markers of V domain of mouse MOPC 21 IgG₁ L chain. The conformational basis of V_L domain markers and content of MOPC 21 V_L‐like immunoglobulins in sera of inbred mouse strains