Human beta-glucosidase: inhibition by sulphates and purification by affinity chromatography on Dextran-sulphate-Sepharose.
The acid beta-glucosidase (D-glucosyl-N-acylsphingosine glucohydrolase, EC 3.2.1.45) from human placenta is inhibited by sulphated macromolecules such as Dextran sulphate or chondroitin sulphate. This inhibition is alleviated by compounds such as crude taurocholate or phospholipids, which are known activators of acid beta-glucosidase. Partially-purified human beta-glucosidase will bind to Dextran sulphate linked to Sepharose 4B and can be eluted with low concentrations of crude sodium taurocholate. This procedure gives a 10-15 fold purification with good yield and has been included in a scheme giving an approx. 4000-fold purification of placental beta-glucosidase. Evidence is presented which suggests that phospholipids bind to beta-glucosidase by both ionic and hydrophobic interactions. The inhibition of enzyme activity caused by sulphated compounds and non-ionic detergents may be attributed to interference with, respectively, the ionic and hydrophobic binding of phospholipid to the enzyme.
D002846:Chromatography, Affinity / Q000379:methods; D016264:Dextran Sulfate; D003911:Dextrans; D005260:Female; D005959:Glucosidases / Q000302:isolation & purification*; D005962:Glucosylceramidase / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D006801:Humans; D008970:Molecular Weight; D017830:Octoxynol; D010743:Phospholipids / Q000378:metabolism; D011092:Polyethylene Glycols; D011247:Pregnancy; D012685:Sepharose; D013431:Sulfates / Q000494:pharmacology; D013656:Taurocholic Acid / Q000378:metabolism
D003911:Dextrans; D010743:Phospholipids; D013431:Sulfates; D011092:Polyethylene Glycols; D013656:Taurocholic Acid; D017830:Octoxynol; D012685:Sepharose; D016264:Dextran Sulfate; D005959:Glucosidases; D005962:Glucosylceramidase