Delipidization of membrane glycoproteins solubilized in acidified chloroform/methanol. Preparation of N-retinylidene opsin in a water soluble form without-detergent

Bovine retina membrane proteins and glycoproteins were insoluble in chloroform/methanol (2:1, v/v) unless the membrane suspension was precipitated with trichloroacetic acid and the organic solvent mixture added to the precipitated membranes. The presence of millimolar amount of trichloroacetic acid...

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Bibliographic Details
Published inBiochemistry international Vol. 11; no. 1; p. 107
Main Authors Curtino, J A, Chiodi, J C
Format Journal Article
LanguageEnglish
Published Australia 01.07.1985
Subjects
Animals
Cattle
Chloroform
Detergents
Eye Proteins - isolation & purification
Glycoproteins - isolation & purification
Hydrogen-Ion Concentration
Membrane Lipids - isolation & purification
Membrane Proteins - isolation & purification
Methanol
Retinal Pigments - isolation & purification
Rod Opsins
Solubility
Spectrophotometry
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Summary:Bovine retina membrane proteins and glycoproteins were insoluble in chloroform/methanol (2:1, v/v) unless the membrane suspension was precipitated with trichloroacetic acid and the organic solvent mixture added to the precipitated membranes. The presence of millimolar amount of trichloroacetic acid in the organic solvent led to the total solubilization of membranes. The glycoproteins precipitated at the interphase after partition of the acidified chloroform/methanol solution with water and were resolubilized from the interphase with chloroform/methanol/water (1:1:0.3, by vol). The solubility properties of the membrane glycoproteins in the acidified organic solvent mixtures allow to remove the bulk of membrane lipids and to recover from the chloroform/methanol/water solution the glycoprotein of rod outer segment membranes, rhodopsin, as protonated N-retinylidene opsin in a water soluble form.
ISSN:0158-5231