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Bacterial expression of human muscarinic receptor fusion proteins and generation of subtype‐specific antisera
Author(s) -
Levey Allan I.,
Stormann Tom M.,
Brann Mark R.
Publication year - 1990
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(90)81440-y
Subject(s) - fusion protein , antiserum , muscarinic acetylcholine receptor , chemistry , receptor , microbiology and biotechnology , biology , antibody , biochemistry , genetics , recombinant dna , gene
A family of five muscarinic acetylcholine receptor genes (m1–m5) encode highly related proteins; however, for methodological reasons it has not been possible to detect the gene product individually. To develop antibody probes specific for the receptor subtypes, unique regions of m1–m5 cDNAs, corresponding to the third cytoplasmic (13) loops, were subcloned into bacterial expression vectors and the fusion proteins expressed in E. coli were used to generate rabbit antisera. These antisera react specifically with the respective fusion proteins on immunoblots and selectively immunoprecipitate each of the native cloned receptors. Since the 13 loops are immunogenic and the epitopes in the cloned receptors are accessible to antibodies, this approach should be valuable for immunological studies of the native receptors.