Identification of an immunodominant region on the I-A β chain using site-directed mutagenesis and DNA-mediated gene transfer

J. M. Buerstedde, Larry R Pease, M. P. Bell, A. E. Nilson, G. Buerstedde, D. Murphy, D. J. McKean

Research output: Contribution to journalArticle

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Abstract

To identify which polymorphic residues determine the allospecific antibody binding sites on A(β) polypeptides, mutant A(β)(k) genes were constructed encoding single or multiple amino acids of the d allele at 14 polymorphic positions in the β1 domain. Cell lines expressing these genes were analyzed by quantitative immunofluorescence using 16 mAbs reactive to A(β)(k) or A(β)(d). Substitution of d allele residues at positions 63 and 65-67 in the A(β)(k) polypeptide resulted in the loss of binding of all A(β)(k)-reactive antibodies and the gain of binding of most A(β)(d)-reactive antibodies. Two A(β)(d)-reactive mAbs bound to the mutant A(β)(k) polypeptide containing d allele-characteristic residue at position 40. In contrast, substitution of the other polymorphic residues in the NH2-terminal and COOH-terminal regions of the β1 domain did not alter antibody binding.

Original languageEnglish (US)
Pages (from-to)473-487
Number of pages15
JournalJournal of Experimental Medicine
Volume167
Issue number2
StatePublished - 1988

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Immunodominant Epitopes
Site-Directed Mutagenesis
Alleles
Peptides
Antibodies
DNA
Antibody Binding Sites
Genes
Fluorescent Antibody Technique
Amino Acids
Cell Line

ASJC Scopus subject areas

  • Immunology

Cite this

Buerstedde, J. M., Pease, L. R., Bell, M. P., Nilson, A. E., Buerstedde, G., Murphy, D., & McKean, D. J. (1988). Identification of an immunodominant region on the I-A β chain using site-directed mutagenesis and DNA-mediated gene transfer. Journal of Experimental Medicine, 167(2), 473-487.

Identification of an immunodominant region on the I-A β chain using site-directed mutagenesis and DNA-mediated gene transfer. / Buerstedde, J. M.; Pease, Larry R; Bell, M. P.; Nilson, A. E.; Buerstedde, G.; Murphy, D.; McKean, D. J.

In: Journal of Experimental Medicine, Vol. 167, No. 2, 1988, p. 473-487.

Research output: Contribution to journalArticle

Buerstedde, JM, Pease, LR, Bell, MP, Nilson, AE, Buerstedde, G, Murphy, D & McKean, DJ 1988, 'Identification of an immunodominant region on the I-A β chain using site-directed mutagenesis and DNA-mediated gene transfer', Journal of Experimental Medicine, vol. 167, no. 2, pp. 473-487.
Buerstedde, J. M. ; Pease, Larry R ; Bell, M. P. ; Nilson, A. E. ; Buerstedde, G. ; Murphy, D. ; McKean, D. J. / Identification of an immunodominant region on the I-A β chain using site-directed mutagenesis and DNA-mediated gene transfer. In: Journal of Experimental Medicine. 1988 ; Vol. 167, No. 2. pp. 473-487.
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AU - Nilson, A. E.

AU - Buerstedde, G.

AU - Murphy, D.

AU - McKean, D. J.

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AB - To identify which polymorphic residues determine the allospecific antibody binding sites on A(β) polypeptides, mutant A(β)(k) genes were constructed encoding single or multiple amino acids of the d allele at 14 polymorphic positions in the β1 domain. Cell lines expressing these genes were analyzed by quantitative immunofluorescence using 16 mAbs reactive to A(β)(k) or A(β)(d). Substitution of d allele residues at positions 63 and 65-67 in the A(β)(k) polypeptide resulted in the loss of binding of all A(β)(k)-reactive antibodies and the gain of binding of most A(β)(d)-reactive antibodies. Two A(β)(d)-reactive mAbs bound to the mutant A(β)(k) polypeptide containing d allele-characteristic residue at position 40. In contrast, substitution of the other polymorphic residues in the NH2-terminal and COOH-terminal regions of the β1 domain did not alter antibody binding.

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