Evidence for an interaction between adducin and Na(+)-K(+)-ATPase: relation to genetic hypertension.
Adducin point mutations are associated with genetic hypertension in Milan hypertensive strain (MHS) rats and in humans. In transfected cells, adducin affects actin cytoskeleton organization and increases the Na(+)-K(+)-pump rate. The present study has investigated whether rat and human adducin polymorphisms differently modulate rat renal Na(+)-K(+)-ATPase in vitro. We report the following. 1) Both rat and human adducins stimulate Na(+)-K(+)-ATPase activity, with apparent affinity in tens of nanomolar concentrations. 2) MHS and Milan normotensive strain (MNS) adducins raise the apparent ATP affinity for Na(+)-K(+)-ATPase. 3) The mechanism of action of adducin appears to involve a selective acceleration of the rate of the conformational change E(2) (K) --> E(1) (Na) or E(2)(K). ATP --> E(1)Na. ATP. 4) Apparent affinities for mutant rat and human adducins are significantly higher than those for wild types. 5) Recombinant human alpha- and beta-adducins stimulate Na(+)-K(+)-ATPase activity, as do the COOH-terminal tails, and the mutant proteins display higher affinities than the wild types. 6) The cytoskeletal protein ankyrin, which is known to bind to Na(+)-K(+)-ATPase, also stimulates enzyme activity, whereas BSA is without effect; the effects of adducin and ankyrin when acting together are not additive. 7) Pig kidney medulla microsomes appear to contain endogenous adducin; in contrast with purified pig kidney Na(+)-K(+)-ATPase, which does not contain adducin, added adducin stimulates the Na(+)-K(+)-ATPase activity of microsomes only about one-half as much as that of purified Na(+)-K(+)-ATPase. Our findings strongly imply the existence of a direct and specific interaction between adducin and Na(+)-K(+)-ATPase in vitro and also suggest the possibility of such an interaction in intact renal membranes.
Ferrandi
M
M
Prassis Research Institute Sigma-Tau, 20019 Settimo Milanese, Italy.
Salardi
S
S
Tripodi
G
G
Barassi
P
P
Rivera
R
R
Manunta
P
P
Goldshleger
R
R
Ferrari
P
P
Bianchi
G
G
Karlish
S J
SJ
D000818:Animals; D017487:Ankyrins / Q000494:pharmacology; D002148:Calmodulin-Binding Proteins / Q000235:genetics / Q000302:isolation & purification / Q000494:pharmacology / Q000502:physiology*; D004912:Erythrocytes / Q000201:enzymology / Q000378:metabolism; D006801:Humans; D006973:Hypertension / Q000235:genetics* / Q000378:metabolism*; D007668:Kidney / Q000201:enzymology / Q000378:metabolism; D009154:Mutation / Q000502:physiology; D051381:Rats; D011919:Rats, Inbred Strains; D011994:Recombinant Proteins / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology*; D000099291:Adducins
D017487:Ankyrins; D002148:Calmodulin-Binding Proteins; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D000254:Sodium-Potassium-Exchanging ATPase; D000099291:Adducins