IgE structure-function relationships defined by sequence directed antibodies induced by synthetic peptides

Michael W. Robertson, Fu-Tong Liu

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Six peptides, representing contiguous amino acid sequences within the Cε{lunate}2, Cε{lunate}3 and Cε{lunate}4 domains ofmurine IgE, were selected for synthesis on the basis of overall hydropathy, degree of homology to both human and rat IgE and, where possible, inclusion of a native cysteine residue. Antibodies were produced against each peptide by immunizing rabbits with peptide-KLH conjugates. Each anti-peptide antiserum exhibited good reactivity with the corresponding immunizing peptide (titer: 10-4 to 10-5) and four of the six antisera exhibited a distinct preferential murine IgE reactivity compared to four other murine immunoglobulin classes (IgG1, IgG2b, IgM and IgA). In addition, one antiserum (anti-ε{lunate} peptide 5), raised against a peptide with 80% homology to human IgE, reacted comparably with both human and murine IgE. Each IgE-reactive antiserum was screened for the ability to stimulate mediator release from IgE-sensitized rat basophilic leukemia (RBL) cells. Two of the four IgE-reactive antisera strongly stimulated 3H-serotonin release (anti-ε{lunate} peptides 4 and 5), one antiserum showed weak activity (anti-ε{lunate} peptide 3) and the remaining anti-peptide serum (anti-ε{lunate} peptide 6), which exhibited the highest anti-IgE reactivity, exhibited no detectable stimulatory activity. Individual anti-peptide antibodies were subsequently tested for the potential to bind to receptor-bound IgE. Anti-ε{lunate} peptide 3 was shown to exhibit the least binding, anti-ε{lunate} peptide 6 showed the highest magnitude of binding while anti-ε{lunate} peptides 4 and 5 exhibited intermediate values. We conclude from this study that sequences defined by ε{lunate}-peptides 4 and 5 are not significantly involved in the receptor binding mechanism whereas ε{lunate} -peptide 3 is likely to be most proximal to the IgE-receptor recognition site of those sequences studied. Finally, we suggest that the ε{lunate}-peptide 6 sequence is in such an orientation in cell-bound IgE that, while it is accessible to external antibody, effective cross-linking of the IgE-receptor complex cannot be achieved through this determinant.

Original languageEnglish (US)
Pages (from-to)103-113
Number of pages11
JournalMolecular Immunology
Volume25
Issue number2
DOIs
StatePublished - 1988

Fingerprint

Immunoglobulin E
Peptides
Antibodies
Immune Sera
IgE Receptors
Immunoglobulin Isotypes
Immunoglobulin A
Cysteine
Immunoglobulin M
Anti-Idiotypic Antibodies
Amino Acid Sequence
Serotonin

ASJC Scopus subject areas

  • Molecular Biology
  • Immunology

Cite this

IgE structure-function relationships defined by sequence directed antibodies induced by synthetic peptides. / Robertson, Michael W.; Liu, Fu-Tong.

In: Molecular Immunology, Vol. 25, No. 2, 1988, p. 103-113.

Research output: Contribution to journalArticle

@article{dd06b11928a8449cbb3541e16b6ceb1e,
title = "IgE structure-function relationships defined by sequence directed antibodies induced by synthetic peptides",
abstract = "Six peptides, representing contiguous amino acid sequences within the Cε{lunate}2, Cε{lunate}3 and Cε{lunate}4 domains ofmurine IgE, were selected for synthesis on the basis of overall hydropathy, degree of homology to both human and rat IgE and, where possible, inclusion of a native cysteine residue. Antibodies were produced against each peptide by immunizing rabbits with peptide-KLH conjugates. Each anti-peptide antiserum exhibited good reactivity with the corresponding immunizing peptide (titer: 10-4 to 10-5) and four of the six antisera exhibited a distinct preferential murine IgE reactivity compared to four other murine immunoglobulin classes (IgG1, IgG2b, IgM and IgA). In addition, one antiserum (anti-ε{lunate} peptide 5), raised against a peptide with 80{\%} homology to human IgE, reacted comparably with both human and murine IgE. Each IgE-reactive antiserum was screened for the ability to stimulate mediator release from IgE-sensitized rat basophilic leukemia (RBL) cells. Two of the four IgE-reactive antisera strongly stimulated 3H-serotonin release (anti-ε{lunate} peptides 4 and 5), one antiserum showed weak activity (anti-ε{lunate} peptide 3) and the remaining anti-peptide serum (anti-ε{lunate} peptide 6), which exhibited the highest anti-IgE reactivity, exhibited no detectable stimulatory activity. Individual anti-peptide antibodies were subsequently tested for the potential to bind to receptor-bound IgE. Anti-ε{lunate} peptide 3 was shown to exhibit the least binding, anti-ε{lunate} peptide 6 showed the highest magnitude of binding while anti-ε{lunate} peptides 4 and 5 exhibited intermediate values. We conclude from this study that sequences defined by ε{lunate}-peptides 4 and 5 are not significantly involved in the receptor binding mechanism whereas ε{lunate} -peptide 3 is likely to be most proximal to the IgE-receptor recognition site of those sequences studied. Finally, we suggest that the ε{lunate}-peptide 6 sequence is in such an orientation in cell-bound IgE that, while it is accessible to external antibody, effective cross-linking of the IgE-receptor complex cannot be achieved through this determinant.",
author = "Robertson, {Michael W.} and Fu-Tong Liu",
year = "1988",
doi = "10.1016/0161-5890(88)90057-0",
language = "English (US)",
volume = "25",
pages = "103--113",
journal = "Molecular Immunology",
issn = "0161-5890",
publisher = "Elsevier Limited",
number = "2",

}

TY - JOUR

T1 - IgE structure-function relationships defined by sequence directed antibodies induced by synthetic peptides

AU - Robertson, Michael W.

AU - Liu, Fu-Tong

PY - 1988

Y1 - 1988

N2 - Six peptides, representing contiguous amino acid sequences within the Cε{lunate}2, Cε{lunate}3 and Cε{lunate}4 domains ofmurine IgE, were selected for synthesis on the basis of overall hydropathy, degree of homology to both human and rat IgE and, where possible, inclusion of a native cysteine residue. Antibodies were produced against each peptide by immunizing rabbits with peptide-KLH conjugates. Each anti-peptide antiserum exhibited good reactivity with the corresponding immunizing peptide (titer: 10-4 to 10-5) and four of the six antisera exhibited a distinct preferential murine IgE reactivity compared to four other murine immunoglobulin classes (IgG1, IgG2b, IgM and IgA). In addition, one antiserum (anti-ε{lunate} peptide 5), raised against a peptide with 80% homology to human IgE, reacted comparably with both human and murine IgE. Each IgE-reactive antiserum was screened for the ability to stimulate mediator release from IgE-sensitized rat basophilic leukemia (RBL) cells. Two of the four IgE-reactive antisera strongly stimulated 3H-serotonin release (anti-ε{lunate} peptides 4 and 5), one antiserum showed weak activity (anti-ε{lunate} peptide 3) and the remaining anti-peptide serum (anti-ε{lunate} peptide 6), which exhibited the highest anti-IgE reactivity, exhibited no detectable stimulatory activity. Individual anti-peptide antibodies were subsequently tested for the potential to bind to receptor-bound IgE. Anti-ε{lunate} peptide 3 was shown to exhibit the least binding, anti-ε{lunate} peptide 6 showed the highest magnitude of binding while anti-ε{lunate} peptides 4 and 5 exhibited intermediate values. We conclude from this study that sequences defined by ε{lunate}-peptides 4 and 5 are not significantly involved in the receptor binding mechanism whereas ε{lunate} -peptide 3 is likely to be most proximal to the IgE-receptor recognition site of those sequences studied. Finally, we suggest that the ε{lunate}-peptide 6 sequence is in such an orientation in cell-bound IgE that, while it is accessible to external antibody, effective cross-linking of the IgE-receptor complex cannot be achieved through this determinant.

AB - Six peptides, representing contiguous amino acid sequences within the Cε{lunate}2, Cε{lunate}3 and Cε{lunate}4 domains ofmurine IgE, were selected for synthesis on the basis of overall hydropathy, degree of homology to both human and rat IgE and, where possible, inclusion of a native cysteine residue. Antibodies were produced against each peptide by immunizing rabbits with peptide-KLH conjugates. Each anti-peptide antiserum exhibited good reactivity with the corresponding immunizing peptide (titer: 10-4 to 10-5) and four of the six antisera exhibited a distinct preferential murine IgE reactivity compared to four other murine immunoglobulin classes (IgG1, IgG2b, IgM and IgA). In addition, one antiserum (anti-ε{lunate} peptide 5), raised against a peptide with 80% homology to human IgE, reacted comparably with both human and murine IgE. Each IgE-reactive antiserum was screened for the ability to stimulate mediator release from IgE-sensitized rat basophilic leukemia (RBL) cells. Two of the four IgE-reactive antisera strongly stimulated 3H-serotonin release (anti-ε{lunate} peptides 4 and 5), one antiserum showed weak activity (anti-ε{lunate} peptide 3) and the remaining anti-peptide serum (anti-ε{lunate} peptide 6), which exhibited the highest anti-IgE reactivity, exhibited no detectable stimulatory activity. Individual anti-peptide antibodies were subsequently tested for the potential to bind to receptor-bound IgE. Anti-ε{lunate} peptide 3 was shown to exhibit the least binding, anti-ε{lunate} peptide 6 showed the highest magnitude of binding while anti-ε{lunate} peptides 4 and 5 exhibited intermediate values. We conclude from this study that sequences defined by ε{lunate}-peptides 4 and 5 are not significantly involved in the receptor binding mechanism whereas ε{lunate} -peptide 3 is likely to be most proximal to the IgE-receptor recognition site of those sequences studied. Finally, we suggest that the ε{lunate}-peptide 6 sequence is in such an orientation in cell-bound IgE that, while it is accessible to external antibody, effective cross-linking of the IgE-receptor complex cannot be achieved through this determinant.

UR - http://www.scopus.com/inward/record.url?scp=0023679870&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023679870&partnerID=8YFLogxK

U2 - 10.1016/0161-5890(88)90057-0

DO - 10.1016/0161-5890(88)90057-0

M3 - Article

C2 - 3374491

AN - SCOPUS:0023679870

VL - 25

SP - 103

EP - 113

JO - Molecular Immunology

JF - Molecular Immunology

SN - 0161-5890

IS - 2

ER -