Membrane anchoring and surface distribution of glycohydrolases of human erythrocyte membranes

Citation
G. Goi et al., Membrane anchoring and surface distribution of glycohydrolases of human erythrocyte membranes, FEBS LETTER, 473(1), 2000, pp. 89-94
Citations number
49
Categorie Soggetti
Biochemistry & Biophysics
Journal title
FEBS LETTERS
ISSN journal
00145793 → ACNP
Volume
473
Issue
1
Year of publication
2000
Pages
89 - 94
Database
ISI
SICI code
0014-5793(20000504)473:1<89:MAASDO>2.0.ZU;2-5
Abstract
The membrane anchoring of the following glycohydrolases of human erythrocyt e plasma membranes was investigated: alpha- and beta-D-glucosidase, alpha- and beta-D-galactosidase, beta-D-glucuronidase, N-acetyl-beta-D-glucosamini dase, alpha-D-mannosidase, and alpha-L-fucosidase. Optimized fluorimetric m ethods for the assay of these enzymes were set up. Treatment of the ghost p reparation with 1.0 mol/l (optimal concentration) NaCl caused release rangi ng from 4.2% of alpha-D-glucosidase to 70% of beta-D-galactosidase; treatme nt with 0.4% (optimal concentration) Triton X-100 liberated 5.1% of beta-D- galactosidase to 89% of alpha-D-glucosidase; treatment with 1.75% (optimal concentration) octylglucoside yielded solubilization from 6.3%, of beta-D-g alactosidase to 85% of alpha-D-glucosidase. Treatment with phosphoinositide -specific phospholipase C caused no liberation of any of the studied glycoh ydrolases. These results are consistent with the notion that the above glyc ohydrolases are differently anchored or associated with the erythrocyte pla sma membrane, and provide the methodological basis for inspecting the occur rence of these enzymes in different membrane microdomains. (C) 2000 Federat ion of European Biochemical Societies.