Brain plasma membrane Na+,K+-ATPase is inhibited by acetylated tubulin.
Membranes from brain tissue contain tubulin that can be isolated as a hydrophobic compound by partitioning into Triton X-114. The hydrophobic behavior of this tubulin is due to the formation of a complex with the alpha-subunit of Na+,K+-ATPase. In the present work we show that the interaction of tubulin with Na+K+-ATPase inhibits the enzyme activity. We found that the magnitude of the inhibition is correlated with: (1) concentration of the acetylated tubulin isoform present in the tubulin preparation used, and (2) amount of acetylated tubulin isoform isolated as a hydrophobic compound. In addition, some compounds involved in the catalytic action of Na+K+-ATPase were assayed to determine their effects on the inhibitory capability of tubulin on this enzyme. The inhibitory effect of tubulin was only slightly decreased by ATP at relatively low nucleotide concentration (0.06 mM). NaCl (1-160 mM) and KCl (0.2-10 mM) showed no effect whereas inorganic phosphate abolished the inhibitory effect of tubulin in a concentration-dependent manner.
Casale
C H
CH
Centro de Investigaciones en Química Biológica de Córdoba (CIQUIBIC), UNC-CONICET, Departamento Química Biológica, Facultad de Ciencias Químicas, Universidad Nacional de Cordoba, Argentina.
Alonso
A D
AD
Barra
H S
HS
D000215:Acylation; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000201:enzymology*; D002384:Catalysis; D002462:Cell Membrane / Q000201:enzymology*; D002847:Chromatography, Agarose; D003902:Detergents / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007700:Kinetics; D017830:Octoxynol / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D011092:Polyethylene Glycols / Q000494:pharmacology; D020033:Protein Isoforms; D051381:Rats; D012965:Sodium Chloride / Q000494:pharmacology; D000254:Sodium-Potassium-Exchanging ATPase / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D014404:Tubulin / Q000737:chemistry / Q000378:metabolism*
D003902:Detergents; D010710:Phosphates; D020033:Protein Isoforms; D014404:Tubulin; D011092:Polyethylene Glycols; D012965:Sodium Chloride; D000255:Adenosine Triphosphate; D017830:Octoxynol; C010615:Nonidet P-40; D000254:Sodium-Potassium-Exchanging ATPase