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The interaction of monovalent cations with the sodium pump of low‐potassium goat erythrocytes
Author(s) -
Cavieres J. D.,
Ellory J. C.
Publication year - 1977
Publication title -
the journal of physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.802
H-Index - 240
eISSN - 1469-7793
pISSN - 0022-3751
DOI - 10.1113/jphysiol.1977.sp012001
Subject(s) - chemistry , sodium , potassium , ouabain , membrane , reaction rate constant , hydrolysis , atp hydrolysis , dissociation (chemistry) , biophysics , atpase , kinetics , enzyme , biochemistry , biology , physics , organic chemistry , quantum mechanics
1. The activation by Na ions and the effect of the anti‐L antibody on the sodium pump of low‐potassium type (LK) erythrocytes, have been studied by measuring ouabain‐sensitive ATPase activity of red cell membranes of LK goats. The experimental data were first corrected for incomplete occupation of the external K sites of the pump, using a saturation function obtained from influx experiments. 2. Double‐reciprocal plots of the corrected rates against Na concentration at various fixed K concentrations, yield a pattern of competitive K inhibition when it is assumed that three equivalent sodium sites take part in the internal activation of LK‐(Na+K)‐ATPase. The dissociation constant of Na at each site ( K m ) lies between 10 and 20 m M and that of K as competitive inhibitor ( K i ), between 1.5 and 4.5 m M . 3. The maximal rate of hydrolysis of LK goat (Na + K)‐ATPase is not different from those usually obtained with the high‐potassium type (HK) red cell enzyme. Then, the low pumping rate of LK erythrocytes in physiological conditions is only reflecting the poor Na affinity, both absolute and relative, at the internal Na sites of their sodium pumps. 4. The stimulation of the ouabain‐sensitive ATPase activity by sensitization of the membranes with anti‐L serum, is mediated by a threefold reduction of the K m / K i ratio at each site. K m decreases by a factor of 10, but there is also a smaller diminution of K i . The maximal rate of hydrolysis, however, is unchanged by the anti‐L treatment. The least‐squares fitting of the pooled data by the rate equation, converges better with less than three and more than two equivalent sodium sites. 5. The affinity sequence at two external K sites of the LK goat erythrocyte sodium pump, determined in the presence of 100 m M external Na, is Rb > K > Cs. It is obtained from the concentration dependence in influx experiments, and is the same as reported for human red cells. 6. Cubic‐root Dixon plots of the corrected ouabain‐sensitive ATPase activity against the concentration of K and its congeners, show the sequence Tl > K > Rb > Na > Cs for the affinities at the internal cation sites of the LK sodium pump. Anti‐L treatment decreases the relative magnitude of Na and Cs selectivities, it being not certain whether a Rb—Na transition then occurs. 7. The results are discussed in terms of possible mechanisms whereby the sodium pump of LK and HK red cells may adjust the properties of their cation sites upon translocation of monovalent cations.