The activator of cerebroside sulphatase. Binding studies with enzyme and substrate demonstrating the detergent function of the activator protein.
1. Sulphatase A (cerebroside sulphatase) (EC 3.1.6.1.) and a 12-fold excess of its physiological activator protein were chromatographed together on Sephadex G-75. The elution buffer was the same as that used in the enzymic degradation of sulphatides. The two proteins were eluted in different peaks indicating that no stable complex formed. 2. Activator protein was incubated with sulphatides under conditions used favouring the sulphatase activity. Incubation solutions were then examined by electrophoresis on a polyacrylamide gel gradient. An one-to-one complex between activator and sulphatides was observed. Half maximal binding occurred with 2.5 nmol of sulphatides together with 1 or 2 nmol of activator in 100 micronl. 3. Cerebrosides as the enzymic degradation products of sulphatides, bind also to the activator protein. A ratio of one-to-one could possibly be obtained at high cerebroside concentrations. The binding to cerebrosides is less specific than that to sulphatides. A 7-fold excess of cerebrosides was necessary for half maximal binding. 4. In a mixture of sulphatides and cerebrosides the formation of the complex with the activator protein is partly inhibited. The total amount of bound lipids changed as the composition of the lipid mixture was varied. In a one-to-one mixture of the two lipids 60% of the total bound lipids are sulphatides and 40% are cerebrosides.
D001665:Binding Sites; D002553:Cerebroside-Sulfatase / Q000378:metabolism*; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D008099:Liver / Q000201:enzymology*; D011485:Protein Binding; D011506:Proteins / Q000302:isolation & purification / Q000502:physiology*; D013429:Sulfatases / Q000378:metabolism*; D013433:Sulfoglycosphingolipids
D011506:Proteins; D013433:Sulfoglycosphingolipids; D013429:Sulfatases; D002553:Cerebroside-Sulfatase