Phospholipid reactivation of the purified plasma membrane ATPase of yeast. 1980

J P Dufour, and A Goffeau

The plasma membrane ATPase of the yeast Schizosaccharomyces pombe solubilized by lysolecithin and purified by centrifugation through a sucrose gradient is essentially inactive. The phospholipid distribution in the sucrose gradient indicates that inactivation of the ATPase may result from the partial delipidation occurring during purification. Taking into account the 100,000 daltons of the ATPase polypeptide, it is concluded that 74 mol of phospholipids are bound per mol of purified ATP monomer. The ATPase so purified is reactivated simply by mixing the enzyme with preformed lipid micelles or vesicles. Lysolecithins reactivate the enzyme at concentrations around the critical micellar concentration. Gel exclusion chromatography indicates that the enzyme binds reversibly to the lysolecithin micelles. On the other hand, lecithins of varying chain length and unsaturation reactivate the enzyme to different extents and with different efficiencies. In addition, from binding studies, it is observed that each saturated lecithin combines equally well with the ATPase. Using other diacylphospholipids no specificity for the polar head group is observed. Moreover, cardiolipin microvesicles is shown to bind all the protein but not to restore the enzyme activity. From lipid-reactivation titration curves. Arrhenius plots, and physical data of the phospholipids, it is concluded that the major parameter which governs the optimal reactivation of ATPase is the ability of the phospholipids to form amphipathic structures (micelles or vesicles) of sufficient fluidity and hydrophobicity. From these results, a coherent description can be provided for the protein-lipid interactions occurring during solubilization, purification, and the lipid reactivation of the yeast plasma membrane ATPase.

UI MeSH Term Description Entries
D007700 Kinetics The rate dynamics in chemical or physical systems.
D008563 Membrane Lipids Lipids, predominantly phospholipids, cholesterol and small amounts of glycolipids found in membranes including cellular and intracellular membranes. These lipids may be arranged in bilayers in the membranes with integral proteins between the layers and peripheral proteins attached to the outside. Membrane lipids are required for active transport, several enzymatic activities and membrane formation. Cell Membrane Lipid,Cell Membrane Lipids,Membrane Lipid,Lipid, Cell Membrane,Lipid, Membrane,Lipids, Cell Membrane,Lipids, Membrane,Membrane Lipid, Cell,Membrane Lipids, Cell
D010743 Phospholipids Lipids containing one or more phosphate groups, particularly those derived from either glycerol (phosphoglycerides see GLYCEROPHOSPHOLIPIDS) or sphingosine (SPHINGOLIPIDS). They are polar lipids that are of great importance for the structure and function of cell membranes and are the most abundant of membrane lipids, although not stored in large amounts in the system. Phosphatides,Phospholipid
D002462 Cell Membrane The lipid- and protein-containing, selectively permeable membrane that surrounds the cytoplasm in prokaryotic and eukaryotic cells. Plasma Membrane,Cytoplasmic Membrane,Cell Membranes,Cytoplasmic Membranes,Membrane, Cell,Membrane, Cytoplasmic,Membrane, Plasma,Membranes, Cell,Membranes, Cytoplasmic,Membranes, Plasma,Plasma Membranes
D004789 Enzyme Activation Conversion of an inactive form of an enzyme to one possessing metabolic activity. It includes 1, activation by ions (activators); 2, activation by cofactors (coenzymes); and 3, conversion of an enzyme precursor (proenzyme or zymogen) to an active enzyme. Activation, Enzyme,Activations, Enzyme,Enzyme Activations
D000251 Adenosine Triphosphatases A group of enzymes which catalyze the hydrolysis of ATP. The hydrolysis reaction is usually coupled with another function such as transporting Ca(2+) across a membrane. These enzymes may be dependent on Ca(2+), Mg(2+), anions, H+, or DNA. ATPases,Adenosinetriphosphatase,ATPase,ATPase, DNA-Dependent,Adenosine Triphosphatase,DNA-Dependent ATPase,DNA-Dependent Adenosinetriphosphatases,ATPase, DNA Dependent,Adenosinetriphosphatases, DNA-Dependent,DNA Dependent ATPase,DNA Dependent Adenosinetriphosphatases,Triphosphatase, Adenosine
D001203 Ascomycota A phylum of fungi which have cross-walls or septa in the mycelium. The perfect state is characterized by the formation of a saclike cell (ascus) containing ascospores. Most pathogenic fungi with a known perfect state belong to this phylum. Ascomycetes,Cochliobolus,Sclerotinia,Ascomycete,Ascomycotas,Sclerotinias
D012568 Schizosaccharomyces A genus of ascomycetous fungi of the family Schizosaccharomycetaceae, order Schizosaccharomycetales. Fission Yeast,Schizosaccharomyces malidevorans,Schizosaccharomyces pombe,Yeast, Fission,S pombe,Fission Yeasts
D013329 Structure-Activity Relationship The relationship between the chemical structure of a compound and its biological or pharmacological activity. Compounds are often classed together because they have structural characteristics in common including shape, size, stereochemical arrangement, and distribution of functional groups. Relationship, Structure-Activity,Relationships, Structure-Activity,Structure Activity Relationship,Structure-Activity Relationships

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