Purification and characterization of the membrane (Na+ + Mg2+)-ATPase from Acholeplasma laidlawii B.
The membrane (Na+ + Mg2+)-ATPase of Acholeplasma laidlawii B has been solubilized with a Brij-58/sodium deoxycholate mixture and purified by a combination of gel filtration and ion-exchange chromatography. The purified, partially delipidated ATPase has a specific activity of 195 mumol Pi/mg protein per h, which could be enhanced by 25% upon the addition of exogenous phospholipids. The kinetic properties of the purified enzyme are similar to those of the native membrane-bound enzyme, suggesting that it has not been substantially altered during the purification procedure. The enzyme is an assembly of five polypeptide species and its kinetic properties suggest that it is dissimilar to other known ATPases.
D000128:Acholeplasma laidlawii / Q000201:enzymology*; D000251:Adenosine Triphosphatases / Q000302:isolation & purification* / Q000378:metabolism; D002462:Cell Membrane / Q000201:enzymology; D003432:Cross-Linking Reagents / Q000494:pharmacology; D007700:Kinetics; D046911:Macromolecular Substances; D008970:Molecular Weight; D012995:Solubility; D013379:Substrate Specificity; D013388:Succinimides / Q000494:pharmacology
D003432:Cross-Linking Reagents; D046911:Macromolecular Substances; D013388:Succinimides; D000251:Adenosine Triphosphatases; C035123:magnesium sodium ATPase; C011240:dithiobis(succinimidylpropionate)