Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies.

S. Paul, D. R. Johnson, R. Massey

Research output: Contribution to journalReview article

4 Citations (Scopus)

Abstract

Autoantibodies purified from humans catalyse the hydrolysis of the neurotransmitter, vasoactive intestinal peptide (VIP). Evidence that the hydrolysis of VIP is due to antibodies includes: the antibody preparations are free of detectable non-immunoglobulin (non-Ig) contamination; the hydrolytic activity is removed by precipitation with anti-human IgG antibody; human B lymphoblastoid cells transformed with Epstein-Barr virus secrete hydrolytic antibodies in culture; the Fab fragments of the antibodies exhibit VIP hydrolysis; and affinity chromatography on immobilized VIP permits purification of specific antibodies with greatly enriched hydrolytic and binding activities. One of the catalytic antibody preparations hydrolyses the Gln-16-Met-17 bond. Studies with synthetic VIP fragments showed that the epitope recognized by this antibody is formed by VIP(15-28). Important binding interactions are contributed by VIP(22-28), a sequence four residues distant from the scissile bond. Antibodies from a second subject hydrolyse six peptide bonds in VIP, clustered between residues 14 and 22. These bonds link amino acids of different charge, size and hydrophobicity, suggesting that the hydrolytic repertoire of the antibodies is considerable. The antibodies do not hydrolyse peptides unrelated in sequence to VIP. Cleavage of several peptide bonds in VIP by polyclonal antibody preparations may be due to several antibodies, each with a unique cleavage specificity. Alternatively, a single antibody may make catalytically productive contact at multiple peptide bonds in the substrate, because of conformational flexibility of VIP or of the antibody active site. Purified light chains from the catalytic antibodies hydrolysed VIP more rapidly than did intact antibodies. The residues constituting the catalytic site of an antibody may be encoded in germline V-region genes or may arise during maturation of the antibody response.

Original languageEnglish (US)
Pages (from-to)156-167; discussion 167-173
JournalCiba Foundation symposium
Volume159
StatePublished - 1991

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Catalytic Antibodies
Epitopes
Anti-Idiotypic Antibodies
Vasoactive Intestinal Peptide
Peptides
Antibodies
Hydrolysis
Catalytic Domain
Immunoglobulin Fab Fragments
Peptide Fragments

ASJC Scopus subject areas

  • General

Cite this

Paul, S., Johnson, D. R., & Massey, R. (1991). Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies. Ciba Foundation symposium, 159, 156-167; discussion 167-173.

Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies. / Paul, S.; Johnson, D. R.; Massey, R.

In: Ciba Foundation symposium, Vol. 159, 1991, p. 156-167; discussion 167-173.

Research output: Contribution to journalReview article

Paul, S, Johnson, DR & Massey, R 1991, 'Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies.', Ciba Foundation symposium, vol. 159, pp. 156-167; discussion 167-173.
Paul, S. ; Johnson, D. R. ; Massey, R. / Binding and multiple hydrolytic sites in epitopes recognized by catalytic anti-peptide antibodies. In: Ciba Foundation symposium. 1991 ; Vol. 159. pp. 156-167; discussion 167-173.
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abstract = "Autoantibodies purified from humans catalyse the hydrolysis of the neurotransmitter, vasoactive intestinal peptide (VIP). Evidence that the hydrolysis of VIP is due to antibodies includes: the antibody preparations are free of detectable non-immunoglobulin (non-Ig) contamination; the hydrolytic activity is removed by precipitation with anti-human IgG antibody; human B lymphoblastoid cells transformed with Epstein-Barr virus secrete hydrolytic antibodies in culture; the Fab fragments of the antibodies exhibit VIP hydrolysis; and affinity chromatography on immobilized VIP permits purification of specific antibodies with greatly enriched hydrolytic and binding activities. One of the catalytic antibody preparations hydrolyses the Gln-16-Met-17 bond. Studies with synthetic VIP fragments showed that the epitope recognized by this antibody is formed by VIP(15-28). Important binding interactions are contributed by VIP(22-28), a sequence four residues distant from the scissile bond. Antibodies from a second subject hydrolyse six peptide bonds in VIP, clustered between residues 14 and 22. These bonds link amino acids of different charge, size and hydrophobicity, suggesting that the hydrolytic repertoire of the antibodies is considerable. The antibodies do not hydrolyse peptides unrelated in sequence to VIP. Cleavage of several peptide bonds in VIP by polyclonal antibody preparations may be due to several antibodies, each with a unique cleavage specificity. Alternatively, a single antibody may make catalytically productive contact at multiple peptide bonds in the substrate, because of conformational flexibility of VIP or of the antibody active site. Purified light chains from the catalytic antibodies hydrolysed VIP more rapidly than did intact antibodies. The residues constituting the catalytic site of an antibody may be encoded in germline V-region genes or may arise during maturation of the antibody response.",
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