TY - JOUR
T1 - Identification of three extended antibodybinding segments in recombinant human muscle acetylcholine receptor α subunitextracellular domain 1-210
AU - Sano, Masato
AU - McCormick, Daniel J.
AU - Talib, Sohel
AU - Griesmann, Guy E.
AU - Lennon, Vanda A.
N1 - Funding Information:
This work was supported by NIH grants NS 15057 (V.A.L.), NS 24694 (D.J.M.) and NS 28608 (S.T.), and by the William K. Warren Foundation.
PY - 1991/10
Y1 - 1991/10
N2 - 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.
AB - 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.
KW - Myasthenia gravis
KW - Nicotinic receptor
KW - Recombinant autoantigen
KW - Synthetic peptides
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U2 - 10.1093/intimm/3.10.983
DO - 10.1093/intimm/3.10.983
M3 - Article
C2 - 1721836
AN - SCOPUS:0026042745
SN - 0953-8178
VL - 3
SP - 983
EP - 989
JO - International Immunology
JF - International Immunology
IS - 10
ER -