Comparison of the ability of phospholipids from rat liver lysosomes to reconstitute glucocerebrosidase

Alakananda Basu, Robert H. Glew, John R. Wherrett, Srebrenka Huterer

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

The in situ lipid activator of rat liver glucocerebrosidase was investigated. Rat liver lysosomes were purified (42.9-fold relative to the crude homogenate) by sequential isopycnic centrifugation in sucrose and metrizamide gradients. Lipids were extracted with chloroform:methanol (2:1) and phospholipids were separated by one-dimensional thinlayer chromatography. The phospholipid content of the lysosome preparation was 0.28 μmol lipid phosphorus/mg protein. Phosphatidylcholine was present as the major non-acidic phospholipid (39.3%). Of the acidic phospholipids, phosphatidylinositol and phosphatidylserine were present in the greatest amounts (12.0 and 19.7%, respectively). The resolved phospholipids were tested separately and in the presence of a heat-stable factor from Gaucher spleen for their ability to reconstitute butanol-delipidated rat liver glucocerebrosidase activity. Alone or in the presence of the heat-stable factor, phosphatidylserine and phosphatidylinositol were the most effective activators of glucocerebrosidase. Bis(monoacylglyceryl) phosphate derived from rat liver tritosomes or rabbit lung macrophages was also effective in reconstituting β-glucosidase activity.

Original languageEnglish
Pages (from-to)464-469
Number of pages6
JournalArchives of Biochemistry and Biophysics
Volume245
Issue number2
DOIs
Publication statusPublished - Mar 1986

    Fingerprint

Cite this