Naslov
Epitope mapping by amino-acid-sequence-specific antibodies reveals that both ends of the alpha subunit of Na+/K(+)-ATPase are located on the cytoplasmic side of the membrane

Autori
Antolović, Roberto ; Bruller, Hans Joachim ; Bunk, Sabine ; Linder, Dietmar ; Schoner, Wilhelm

Izvornik
European journal of biochemistry (0014-2956) 199 (1991); 195-202

Vrsta, podvrsta i kategorija rada
Radovi u časopisima, članak, znanstveni

Ključne riječi
sequence specific antibodies ; Na/K-ATPase

Sažetak
Right-side-out vesicles of pig kidney microsomes and amino-acid-sequence-specific antibodies were used to probe the sidedness of the C-terminus and the N- terminus of the catalytic alpha subunit of Na+/K(+)- ATPase. Polyclonal antibodies were raised in rabbits against the peptide corresponding to the N-terminal sequence GRDKYEPAAVSE (peptide 1- 12) and against peptides corresponding to the C- terminal sequences IFVYDEVRKLIIRRR (peptide 991- 1005) and RPGGWVEKETYY (peptide 1005-1016). These antibodies were purified by affinity chromatography on the respective peptide- Sepharose columns. Moreover, antibodies against the N- terminal dodecapeptide GRDKYEPAAVSE were obtained by affinity purification from heteroclonal antibodies against the alpha subunit of pork kidney Na+/K(+)- ATPase. These antibodies reacted with native as well as SDS-denaturated Na+/K(+)- ATPase. When the antibodies were used to probe the sidedness of the sequences in right-side-out vesicles of pig kidney microsomes, the N-terminal peptide 1-12 as well as the C-terminal peptides 991-1005 and 1005-1016 were found on the cytosolic side. Concanavalin A, however, which interacts with the beta subunit, a glycoprotein, reacted with the outside of right-side- out vesicles.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

POVEZANOST RADA