The Attachment of Heinz Bodies to the Red Cell Membrane

S ummary . The binding between Heinz bodies or heat-precipitated haemoglobin and the red cell membrane was investigated. Ghosts containing Heinz bodies prepared from red cells containing Hb Christchurch and ghosts bound to heat-precipitated Hb A were examined by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulphate. Using this technique, which separated the membrane and Heinz body proteins according to size, no protein bands corresponding to derivatives of haemoglobin covalently bound to a ghost protein were detected. Binding between red cell ghosts and heat-precipitated Hb A was also studied by density gradient ultracentrifugation, which separated ghosts bound to haemoglobin from pure ghosts and pure heat precipitate. Sulphydryl reducing and blocking agents were found to have no effect on the attachment of the insoluble haemoglobin to the ghosts, and reduction of electrostatic interactions in 2 m -NaCl was also ineffective.
It is concluded that covalent bonding, and in particular disulphide bonding, is not involved in the attachment of Heinz bodics to the red cell membrane. Hydrophobic bonding is suggested as the likely alternative.