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STUDIES OF THE MECHANISM OF CATION TRANSPORT
Author(s) -
Charnock JS,
Rosenthal AS,
Post RL
Publication year - 1963
Publication title -
australian journal of experimental biology and medical science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.999
H-Index - 104
eISSN - 1440-1711
pISSN - 0004-945X
DOI - 10.1038/icb.1963.56
Subject(s) - chemistry , dephosphorylation , ouabain , sodium , potassium , strophanthin , adenosine triphosphate , phosphorylation , phosphatase , inorganic chemistry , biophysics , biochemistry , organic chemistry , biology
SUMMARY Membrane‐bound cation stimulated adenosine‐triphosphatase is thought to he closely associated with active ion transport phenomena. As this enzyme system requires the combination of both sodium and potassium ions for full activation its mechanism of action has been investigated under these conditions. Using adenosine triphosphate labelled with P 32 only in the terminal position as substrate for the reaction we have been able to show that ATP 32 is hydrolysed to adenosine diphosphate and radioactive orthophosphate via a phosphorylated intermediate. Sodium but not potassium ions are required for the formation of this intermediate and adenosine diphosphate; potassium ions are subsequently required for dephosphorylation of the intermediate complex with the release of P 32 orthophosphate — that is, both sodium and potassium ions arr required for the turnover of the intermediate in a two step reaction. The cardiac glycoside ouabain (Strophanthin‐G) is without effect upon the formation of this intermediate complex, but can completely inhibit the K + — requiring dephosphorylation reaction. Preliminary experiments indicate that sodium ions are bound to the enzyme system during the phosphorylation reaction and are released when this complex is broken down. Solvent extraction procedures designed to remove phospholipid components did not decrease the specific activity of the phosphorylated intermediate which suggests that it is phosphoprotein in nature. These results support the belief that Na + + K + activated ATPase participates in active cation transport in a physical as well as chemical sense, and a possible biochemical mechanism of active cation transport is suggested.