PMID: 6105871Apr 15, 1980Paper

Reconstitution of the Ca2+-transport system of human erythrocytes

The Biochemical Journal
K GietzenH U Wolf

Abstract

The (Ca2+ + Mg2+)-dependent ATPase of human erythrocyte 'ghosts' was solubilized and reconstituted to form membranous vesicles capable of energized Ca2+ accumulation. The erythrocyte 'ghosts' for this purpose were prepared by using isoosmotic freeze-haemolysis in the presence of Tween 20 and proteinase inhibitors to stabilize the preparation. The reconstitution procedure is similar to that developed by Meissner & Fleischer [(1974) J. Biol. Chem. 249, 302-309] for skeletal-muscle sarcoplasmic-reticulum in that: (1) deoxycholate is used for the solubilization of the membrane; (2) controlled dialysis at near room temperature, rather than 0 degree C, is required in order to obtain a functional preparation capable of Ca2+ accumulation; and (3) membrane vesicles can be reassembled with protein/lipid ratio (approx. 60% protein and 40% lipid) similar to that of the original membrane.

Citations

Jan 1, 1982·Annals of the New York Academy of Sciences·B SarkadiG Gárdos
Mar 17, 1986·European Journal of Biochemistry·K MartinekI V Berezin
Feb 1, 1994·In Vitro Cellular & Developmental Biology. Animal·S RauthT K Das Gupta
Nov 1, 1982·Hepatology : Official Journal of the American Association for the Study of Liver Diseases·S LotersztajnF Pecker
Nov 16, 2007·American Journal of Physiology. Cell Physiology·Renate J ScheibePetra Wetzel
May 15, 1990·Biochimica Et Biophysica Acta·A Villalobo
Feb 1, 1985·Archives of Biochemistry and Biophysics·D R Nelson, D J Hanahan
Sep 15, 1988·Biochemical and Biophysical Research Communications·S C KingS M Goldin

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