Structure of epitopes recognized by the antibodies to alpha(181-192) peptides of neuronal nicotinic acetylcholine receptors: extrapolation to the structure of acetylcholine-binding domain.
Using the alpha(181-192) peptides of neuronal nicotinic acetylcholine receptor (nAChR) and Ala-substituted peptide analogues, amino acid residues critical for specific monoclonal antibody (mAb) binding were identified. By means of 2D nuclear magnetic resonance (2D-NMR) analysis followed by molecular modeling, it was found that mAb binding resulted in stabilization of the free alpha3(181-192) peptide flexible conformation yielding an extended structure with residues 6-11 of the peptide being in direct contact with the Ab. Since the Ab binds the native AChR as well, it is suggested that the corresponding fragment of AChR alpha3 subunit is exposed to solution and also appears in extended conformation.
Skok
M
M
Department of Molecular Immunology, Palladin Institute of Biochemistry, 9, Leontovicha str., 01030, Kiev, Ukraine. skok@biochem.kiev.ua
Lykhmus
E
E
Bobrovnik
S
S
Tzartos
S
S
Tsouloufis
T
T
Vanderesse
R
R
Coutrot
F
F
Thong Cung
M
M
Marraud
M
M
Krikorian
D
D
Sakarellos-Daitsiotis
M
M
D000409:Alanine; D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D000918:Antibody Specificity; D001665:Binding Sites / Q000276:immunology; D000939:Epitopes / Q000276:immunology; D009682:Magnetic Resonance Spectroscopy; D008958:Models, Molecular; D009474:Neurons / Q000737:chemistry; D010446:Peptide Fragments / Q000138:chemical synthesis / Q000276:immunology; D017434:Protein Structure, Tertiary; D011978:Receptors, Nicotinic / Q000737:chemistry* / Q000276:immunology*
D000409:Alanine; D000911:Antibodies, Monoclonal; D000939:Epitopes; D010446:Peptide Fragments; D011978:Receptors, Nicotinic