Identification of three extended antibodybinding segments in recombinant human muscle acetylcholine receptor α subunitextracellular domain 1-210

Masato Sano, Daniel J. McCormick, Sohel Talib, Guy E. Griesmann, Vanda A. Lennon

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

The duplicated α subunits account for 40% of the total protein of the nicotinic acetylcholine receptor of muscle, and are implicated as targets for pathogenic autoantibodies in theneuromuscular disease myasthenia gravis (MG). This study reports some of the specificities ofantibodies induced by a myasthenogenic recombinant protein (rHa1 -210) corresponding to theproposed extracellular domain of the a subunit of human acetylcholine receptor, residues 1-210.Antisera produced by immunizing rats, rabbits, and mice were tested with a panel of overlappingsynthetic peptides (each 16 amino acids) comprising residues 1 -216 of the human a subunit. IgG antibodies produced in all three species bound only to peptides that were clustered in three segments: Segment I (residues 9-24); segment II (57-96 in rats, 57-88 in rabbits, and57-80 in mice); and segment III (137-184 in rats, 145-184 in rabbits and mice). Monoclonalantibodies were produced by 41 independent hybridomas derived from three rats immunized withrHa1 -210; 12 reacted only with the recombinant or native protein, and 29 reacted additionallywith peptides in segments II or III. Four mAbs bound to native human receptor; of these, threebound to peptides 57-72/65-80, 81 -96, or 153-168, and one lacked peptide-binding activity.Lack of mAb reactivity with rat receptor precluded correlation of peptide reactivity withmyasthenogenicity. Nevertheless, the data indicate that the human acetylcholine receptor’sa subunit contains multiple sites in its extracellular domain that are potentially stimulatory forВ cells.

Original languageEnglish (US)
Pages (from-to)983-989
Number of pages7
JournalInternational Immunology
Volume3
Issue number10
DOIs
StatePublished - Oct 1991

Keywords

  • Myasthenia gravis
  • Nicotinic receptor
  • Recombinant autoantigen
  • Synthetic peptides

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology

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