Electron spin resonance (ESR) studies of skeletal protein interactions in human erythrocyte membranes exposed to polyanions and in membranes prepared from inositol hexaphosphate (IHP)-incorporated low-affinity erythrocytes.

Previous biophysical investigations, including those from our laboratories, have reported that polyphosphates weaken RBC membrane skeletal protein-protein interactions and decrease hemoglobin affinity for oxygen. We have additionally demonstrated that low-affinity intact RBC's may be produced by inositol hexaphosphate (IHP) incorporation via an osmotic pulse method. In the present electron spin resonance (ESR) study, IHP was shown to cause a concentration-dependent increase in the segmental motion of ghost membrane skeletal proteins, but no alterations in spin-labeled terminal sialic acid. Pyrophosphate and inositol hexasulfate were significantly less effective in altering the physical state of skeletal proteins than was IHP. Additional ESR studies of both the interaction of IHP with membrane skeletal proteins in the presence of hemoglobin and of membranes obtained from osmotic pulse-treated intact cells were performed. The results of all these studies are discussed in terms of previous biophysical investigations of the effects of polyphosphates on membranes and of possible molecular events that occur during the osmotic pulse procedure.