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Topological analysis of the ATP‐gated ionotrophic P2X 2 receptor subunit
Author(s) -
Torres Gonzalo E,
Egan Terrance M,
Voigt Mark M
Publication year - 1998
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/s0014-5793(98)00179-3
Subject(s) - transmembrane domain , transmembrane protein , hek 293 cells , protein subunit , topology (electrical circuits) , membrane topology , extracellular , amino acid , mutagenesis , receptor , cytoplasm , chemistry , biology , microbiology and biotechnology , biochemistry , mutation , gene , mathematics , combinatorics
We investigated the transmembrane topology of the P2X 2 receptor subunit expressed in HEK 293 cells. Initial studies using two P2X subunits expressed in tandem indicated that the amino‐ and carboxy‐termini are on the same side of the membrane. Immunofluorescence studies showed the cytoplasmic orientation of the amino‐ and carboxy‐termini. Finally, N ‐glycosylation scanning mutagenesis revealed that reporter sites inserted into the central loop, but not those in the amino‐ or carboxy‐terminal regions, were glycosylated, thus suggesting an extracellular placement for that domain. Our results support a two‐transmembrane arrangement for P2X receptors with intracellular amino‐ and carboxy‐termini.