Up-regulation of sodium pump activity in Xenopus laevis oocytes by expression of heterologous beta 1 subunits of the sodium pump.
Recent evidence suggests that the beta subunit of the Na+ pump is essential for the alpha subunit to express catalytic activity and for assembly of the holoenzyme in the plasma membrane. We report here that injection into Xenopus laevis oocytes of cRNAs specific for beta 1 subunit isoforms of the Na+ pump of four species (Torpedo californica, chicken, mouse and rat) causes a time-dependent increase in the number of ouabain-binding sites, both in the plasma membrane and in internal membranes. Expression of the beta 1 subunit of the Na+ pump of mouse and rat in the oocytes could be substantiated by immunoprecipitation using a polyclonal antiserum against the mouse beta 1 subunit. Scatchard analysis in permeabilized cells disclosed that the affinity for ouabain is unchanged after expression of each of the beta 1 subunits. A proportional increase in ouabain-sensitive 86Rb+ uptake indicates that the additionally expressed ouabain-binding sites on the cell surface represent functional Na+ pumps. The findings support the concept of Geering. Theulaz, Verrey, Häuptle & Rossier [(1989) Am. J. Physiol. 257, C851-C858] that beta 1 subunits expressed in oocytes associate with an excess of endogenous alpha subunits of the Na+ pump to form a hybrid enzyme. In addition, all of the beta 1 isoforms investigated in the present study were also capable of combining with the co-expressed alpha 1 subunit of the Torpedo Na+ pump to produce a functional enzyme. Injection of cRNA encoding for the Torpedo alpha 1 subunit alone had no effect on the ouabain-binding capacity of the surface and intracellular membranes of the oocyte.
Schmalzing
G
G
Max-Planck-Institut für Biophysik, Frankfurt/Main, Germany.
Gloor
S
S
Omay
H
H
Kröner
S
S
Appelhans
H
H
Schwarz
W
W
D000818:Animals; D002645:Chickens; D005260:Female; D015870:Gene Expression; D007163:Immunosorbent Techniques; D007700:Kinetics; D046911:Macromolecular Substances; D008297:Male; D009865:Oocytes / Q000378:metabolism*; D010042:Ouabain / Q000378:metabolism; D055503:Protein Multimerization; D012313:RNA / Q000235:genetics; D018075:RNA, Complementary; D051381:Rats; D012414:Rubidium Radioisotopes; D012964:Sodium / Q000378:metabolism; D000254:Sodium-Potassium-Exchanging ATPase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D013045:Species Specificity; D014101:Torpedo; D014162:Transfection; D014982:Xenopus laevis
D046911:Macromolecular Substances; D018075:RNA, Complementary; D012414:Rubidium Radioisotopes; D010042:Ouabain; D012313:RNA; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase