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Membrane anchoring and surface distribution of glycohydrolases of human erythrocyte membranes
Author(s) -
Goi Giancarlo,
Bairati Chiara,
Massaccesi Luca,
Lovagnini Augusto,
Lombardo Adriana,
Tettamanti Guido
Publication year - 2000
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(00)01504-0
Subject(s) - membrane , enzyme , beta (programming language) , alpha (finance) , chemistry , erythrocyte membrane , chromatography , biochemistry , medicine , construct validity , nursing , computer science , patient satisfaction , programming language
The membrane anchoring of the following glycohydrolases of human erythrocyte plasma membranes was investigated: α‐ and β‐ D ‐glucosidase, α‐ and β‐ D ‐galactosidase, β‐ D ‐glucuronidase, N ‐acetyl‐β‐ D ‐glucosaminidase, α‐ D ‐mannosidase, and α‐ L ‐fucosidase. Optimized fluorimetric methods for the assay of these enzymes were set up. Treatment of the ghost preparation with 1.0 mol/l (optimal concentration) NaCl caused release ranging from 4.2% of α‐ D ‐glucosidase to 70% of β‐ D ‐galactosidase; treatment with 0.4% (optimal concentration) Triton X‐100 liberated 5.1% of β‐ D ‐galactosidase to 89% of α‐ D ‐glucosidase; treatment with 1.75% (optimal concentration) octylglucoside yielded solubilization from 6.3% of β‐ D ‐galactosidase to 85% of α‐ D ‐glucosidase. Treatment with phosphoinositide‐specific phospholipase C caused no liberation of any of the studied glycohydrolases. These results are consistent with the notion that the above glycohydrolases are differently anchored or associated with the erythrocyte plasma membrane, and provide the methodological basis for inspecting the occurrence of these enzymes in different membrane microdomains.