Identification of a peptide that protects the human acetylcholine receptor against antigenic modulation.
mAb 192 is a rat monoclonal antibody with very high affinity for the major immunogenic region (MIR) of the human muscle acetylcholine receptor (AChR). An epitope mimic of this antibody was selected from a phage display peptide library screened with mAb 192. The peptide-presenting phage has been shown to specifically bind to solid phase mAb 192 with an equilibrium dissociation constant (K(d)) of 8.45x10(-9) M, as directly measured with surface plasmon resonance. This value represents the avidity of the interaction between selected phage and mAb 192. A synthetic version of this peptide QPSPYNGWRMEI, referred to as MG15, binds to its selecting antibody and blocks the interaction of mAb 192 with human AChR. Peptide MG15 was able to protect acetylcholine receptors on human RD cells from antibody-mediated down-modulation. The negative charge of glutamic acid plays a important role in antibody binding. Replacement of the glutamic acid with an alanine completely abolishes the inhibitory activity.
Luo
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GX
La Jolla Pharmaceutical Company, 6455 Nancy Ridge Drive, San Diego, CA 92121, USA.
Victor
K
K
Chong
K
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McNeeley
P
P
Ramirez
D
D
Preclaro
J
J
Linnik
M D
MD
Campbell
M A
MA
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D000941:Antigens / Q000235:genetics; D001483:Base Sequence; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D007700:Kinetics; D019151:Peptide Library; D010455:Peptides / Q000235:genetics / Q000276:immunology* / Q000494:pharmacology; D051381:Rats; D011950:Receptors, Cholinergic / Q000235:genetics / Q000276:immunology*; D020349:Surface Plasmon Resonance
D000911:Antibodies, Monoclonal; D000941:Antigens; D017931:DNA Primers; D019151:Peptide Library; D010455:Peptides; D011950:Receptors, Cholinergic