Kinetics of the Reaction Between DNP-Insulin and Homologous Antibody

Lee, F. H.; Froese, Arnold

doi:10.3109/08820137309022827

Cited by 4 publications

(1 citation statement)

References 12 publications

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“…The increased association rate observed with the hexyl chain (Table 4) may reflect the greater conformational freedom of the Dnp group. The effect of flexibility is clearly illustrated in the association between [Dnp-Lys29]insulin and its homologous anti-Dnp antibody (Lee & Froese, 1973). In this immunogen the Dnp group is attached to a flexible portion of the polypeptide chain (Blundell et al, 1972) and the asiociation rate constant is approx.…”

Section: Discussionmentioning

confidence: 99%

The accessibility of protein-bound dinitrophenyl groups to univalent fragments of anti-dinitrophenyl antibody

Knight¹,

Green²

1976

Biochemical Journal

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A series of N-(N-dinitrophenylaminoalkyl)maleimides were sythesized with alkyl-chain lengths of two, four and six carbon atoms. When these compounds reacted with the thiol group of mercaptalbumin, the tryptophan fluorescence of the protein was quenched. This change in fluorescence was used to determine the rate of reaction of the Dnp (dinitrophenyl)-maleimides with mercaptalbumin. The second-order rate constants were similar to those observed in reactions between low-molecular-weight thiol compounds and maleimides. When N-(N-Dnp-aminoalkyl)succinimidomercaptalbumins were added to univalent fragments of anti-Dnp antibody the antibody fluorescence was quenched. Florescence-quenching titrations showed that the protein-bound Dnp groups were fully available to the antibody even when the alkyl chain was short. The apparent dissociation constants were significantly greater than that of the interaction between anti-Dnp antibody and the free hapten, 6-(N-Dnp)-aminohexanoate. The antibody fluorescence was quenched efficienty by [dnp-Lys41]ribonuclease A, also with an increased dissociation constant. It could be concluded from the increase in dissociation constant that the Dnp group spent no more than 0.1% of its time in the dissociated state, available to antibody. The second-order rate constants for the association between the Dnp-mercaptablumins and the antibody were determined and were similar in magnitude to those observed in other interactions between protein and anti-protein antibody.

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Section: Discussionmentioning

confidence: 99%

The accessibility of protein-bound dinitrophenyl groups to univalent fragments of anti-dinitrophenyl antibody

Knight¹,

Green²

1976

Biochemical Journal

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Ir gene control of carrier recognition. I. Immunogenicity of bovine insulin derivatives

Keck

1975

Eur J Immunol

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2,4‐Dinitrophenyl‐lysin B29 ‐bovine insulin was used as immunogen to investigate the carrier function of insulin in the induction of immune response. Experiments with backcross mice suggest that this is controlled by autosomal Ir genes. The response pattern shown by congenic mice demonstrate the linkage of these genes to the H‐2 complex. With a standard dose of 20 μg per mouse, strains with the H‐2 haplotypes d,b,v respond to this carrier. All other strains tested, with the H‐2 haplotypes a,k,u,r,s,q, are nonresponders. The two Ir genes, Ir‐insulinBov (H‐2d) and Ir‐insulinBov (H‐2b), which are associated with H‐2d and H‐2b respectively, can be distinguished from one another on a functional basis. Only H‐2d mice respond to an appreciable extent when B. pertussis organisms are the adjuvant. With complete Freund's adjuvant, H‐2d and H‐2b mice give high antibody titers, but with low doses of antigen only H‐2d mice respond. In contrast to H‐2d mice, H‐2b mice do not respond to trifluoresceinthiocarbamyl insulin, suggesting that the dimeric insulin might be the immunogenic form in these mice. Furthermore, as has been shown earlier (Nature, 1975. 254: 78.), the A‐chain loop is needed as a carrier determinant only in H‐2b mice. Experiments with recombinant strains localize Ir‐insulinBov (H‐2b) to the left of Ir‐1B, probably in Ir‐1A. H‐2k mice are strict nonresponders, even when other proteins such as bovine IgG, methylated bovine serum albumin or rabbit IgG are provided as carriers complexed with insulin. However, if proinsulin is used as the antigen, anti‐insulin antibody can easily be raised in these strains.

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Kinetics of Antibody-Hapten Interactions

Pecht¹,

Lancet²

1977

Molecular Biology Biochemistry and Biophysics

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Kinetics of the Reaction Between DNP-Insulin and Homologous Antibody

Cited by 4 publications

References 12 publications

The accessibility of protein-bound dinitrophenyl groups to univalent fragments of anti-dinitrophenyl antibody

The accessibility of protein-bound dinitrophenyl groups to univalent fragments of anti-dinitrophenyl antibody

Ir gene control of carrier recognition. I. Immunogenicity of bovine insulin derivatives

Kinetics of Antibody-Hapten Interactions

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