In situ assembly states of (Na+,K+)-pump ATPase in human erythrocytes. Radiation target size analyses.
The in situ assembly state of the (Na+,K+)-pump ATPase of human erythrocytes was studied by applying the classical target theory to radiation inactivation data of the ouabain-sensitive sodium efflux and ATP hydrolysis. Erythrocytes and their extensively washed white ghosts were irradiated at -45 to -50 degrees C with an increasing dose of 1.5-MeV electron beam, and after thawing, the Na+-pump flux and/or enzyme activities were assayed. Each activity measured was reduced as a simple exponential function of radiation dose, from which a radiation sensitive mass (target size) was calculated. When intact cells were used, the target sizes for the pump and for the ATPase activities were equal and approximately 620,000 daltons. The target size for the ATPase activity was reduced to approximately 320,000 daltons if the cells were pretreated with digitoxigenin. When ghosts were used, the target size for the ATPase activity was again approximately 320,000 daltons. Our target size measurements together with other information available in literature suggest that (Na+,K+)-pump ATPase may exist in human erythrocytes either as a tetramer of alpha beta or as a dimer of alpha beta in tight association with other protein mass, probably certain glycolytic enzymes, and that this tetrameric or heterocomplex association is dissociable by digitoxigenin treatment or by extensive wash during ghost preparation.
J
J
Hah
J M
JM
Goldinger
C Y
CY
Jung
D004073:Digitoxigenin / Q000494:pharmacology; D004910:Erythrocyte Membrane / Q000201:enzymology; D004912:Erythrocytes / Q000187:drug effects / Q000201:enzymology*; D006801:Humans; D007473:Ion Channels / Q000378:metabolism*; D046911:Macromolecular Substances; D008970:Molecular Weight; D010042:Ouabain / Q000494:pharmacology; D012964:Sodium / Q000097:blood; D000254:Sodium-Potassium-Exchanging ATPase / Q000097:blood* / Q000528:radiation effects
D007473:Ion Channels; D046911:Macromolecular Substances; D004073:Digitoxigenin; D010042:Ouabain; D012964:Sodium; D000254:Sodium-Potassium-Exchanging ATPase