Sickle hemoglobin is more fusogenic than normal hemoglobin at physiological pH and ionic strength conditions.
We used electron microscopy, quasi-elastic light scattering and static light scattering to show that human hemoglobin (Hb) interacts with bovine brain phosphatidylserine lipid vesicles and promotes vesicle fusion in an isotonic buffer at pH 7.4. The fusogenic properties of Hb were observed in both small unilamellar vesicles (SUVs) and large unilamellar vesicles (LUVs). A simple turbidity measurement method was used to follow increases in vesicle size (scattering diameter) as a function of time. For the first 3 h, upon incubation with oxygenated Hb, the scattering diameters of vesicles increased at a rate of 7.8 nm/h for LUVs. Continuous incubation with Hb led to complicated vesicle fusion, probably due to the oxidation products of Hb and lipid molecules. In the absence of both Hb and lipid oxidation, using Hb liganded with carbon monoxide, we obtained, for the entire 20 h incubation period, a fusion rate of 2.9 nm/h for LUVs. We also studied interactions between sickle Hb and vesicles under the same conditions and found that the vesicle fusion rates for sickle Hb were about 2 times faster than those for normal Hb. These results showed that sickle Hb exhibited more extensive interactions with lipid bilayer than normal Hb at physiological pH and ionic strength conditions, and provide insights toward understanding the molecular mechanisms in sickle cell abnormalities.
LaBrake
C C
CC
Department of Chemistry, Loyola University of Chicago, Chicago, IL 60626, USA.
Fung
L W
LW
D002263:Carboxyhemoglobin; D006441:Hemoglobin A; D006451:Hemoglobin, Sickle; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008561:Membrane Fusion; D008567:Membranes, Artificial; D009994:Osmolar Concentration; D010084:Oxidation-Reduction; D010108:Oxyhemoglobins; D010316:Particle Size; D010718:Phosphatidylserines
D006451:Hemoglobin, Sickle; D008567:Membranes, Artificial; D010108:Oxyhemoglobins; D010718:Phosphatidylserines; D006441:Hemoglobin A; D002263:Carboxyhemoglobin