Monoclonal antibodies against the acetylcholine receptor γ‐subunit as site specific probes for receptor tyrosine phosphorylation

FEBS Letters

Tzartos, Elisabeth¹  Tzartos, John S.¹  Tzartos, Socrates J.¹ 

[1] Department of Biochemistry, Hellenic Pasteur Institute, 127, Vas. Sofias Ave., Athens 11521, Greece

关键词: Acetylcholine receptor;    Monoclonal antibody;    Tyrosine phosphorylation;    Epitope mapping;    Synthetic peptide;    AChR;    acetylcholine receptor;    mAb;    monoclonal antibody;    PBS;    145 mM NaCl/7.5 mM Na2HP04/2.5 mM NaH2PO4;    pH 7.4 buffer;    ELISA;    enzyme-linked immunosorbent assay;    SDS;    sodium dodecyl sulphate;    SDS-PAGE;    SDS-polyacrylamide gel electrophoresis;   

DOI  :  10.1016/0014-5793(95)00316-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Tyrosine phosphorylation of the nicotinic acetylcholine receptor (AChR) may be involved in AChR desensitization and clustering. Torpedo AChR γ-subunit is phosphorylated at Tyr³⁶⁵. Using overlapping synthetic peptides, we have precisely mapped the epitopes of five anti-γ-subunit monoclonal antibodies (mAbs) and found that the epitope(s) for the mAbs 154, 165 and 168 (γ365–370) all contain Tyr³⁶⁵. mAb 168 is a known blocker of AChR channel function. Using peptide analogues, Tyr³⁶⁵. was found to be indispensable for mAb165 binding; furthermore its binding was selectively inhibited by in vitro AChR tyrosine phosphorylation. The possible connection between γ-subunit phosphorylation and regulation of AChR function and the proven usefulness of these mAbs as tools should facilitate functional studies of AChR γ-subunit phosphorylation.

【 授权许可】

Unknown