IS HEMOGLOBIN AN ESSENTIAL STRUCTURAL COMPONENT OF HUMAN ERYTHROCYTE MEMBRANES?*

Abstract

This investigation of human red cell membranes was undertaken in order to determine whether the presence of hemoglobin in the membrane is es- sential for maintenance of the membrane's struc- tural integrity, which has been evaluated by stud- ies of the shape, lipid content and composition, protein content, osmotic responsiveness, and certain enzymic activities of erythrocyte ghosts from which virtually all of the cellular hemoglobin had been removed.Many studies have been done with red cell "ghosts," or posthemolytic residues, because such preparations offer the possibility of analyzing certain physiological, biochemical, electron micro- scopic, and immunologic properties attributable to the membranes themselves, apart from the intracel- lular contents.Representative techniques for preparation of red cell ghosts include hemolysis in distilled water, the effects of which were stud- ied first by Bernstein and his associates (1), the freeze-thaw method described by Jorpes (2), the use of chemical lysins such as digitonin and sa- ponin (3) and the use of 0.1%o sodium chloride saturated with CO2 (4), both as described by Ponder, and the method of successive osmotic hemolysis proposed by Hillier and Hoffman (5).Recently, Danon, Nevo, and Marikovsky (6) have described a modification of the technique of Hil- lier and Hoffman which they have termed "gradual osmotic lysis."This method provides a high yield of ghosts that bear a remarkable resemblance

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Affiliated Institutions

• University of Rochester US

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