Membrane interactions of hemoglobin variants, HbA, HbE, HbF and globin subunits of HbA: Effects of aminophospholipids and cholesterol |
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Authors: | Poppy Datta Amit Chakrabarty |
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Affiliation: | a Biophysics Division, Kolkata 700064, India b Structural Genomics Section, Saha Institute of Nuclear Physics, 1/AF, Bidhannagar, Kolkata 700064, India c Thalassemia Foundation, Kolkata 700064, India |
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Abstract: | The interaction of hemoglobin with phospholipid bilayer vesicles (liposomes) has been analyzed in several studies to better understand membrane-protein interactions. However, not much is known on hemoglobin interactions with the aminophospholipids, predominantly localized in the inner leaflet of erythrocytes, e.g., phosphatidylserine (PS), phosphatidylethanolamine (PE) in membranes containing phosphatidylcholine (PC). Effects of cholesterol, largely abundant in erythrocytes, have also not been studied in great details in earlier studies. This work therefore describes the study of the interactions of different hemoglobin variants HbA, HbE and HbF and the globin subunits of HbA with the two aminophospholipids in the presence and absence of cholesterol. Absorption measurements indicate preferential oxidative interaction of HbE and alpha-globin subunit with unilamellar vesicles containing PE and PS compared to normal HbA. Cholesterol was found to stabilize such oxidative interactions in membranes containing both the aminophospholipids. HbE and alpha-globin subunits were also found to induce greater leakage of membrane entrapped carboxyfluorescein (CF) using fluorescence measurements. HbE was found to induce fusion of membrane vesicles containing cholesterol and PE when observed under electron microscope. Taken together, these findings might be helpful in understanding the oxidative stress-related mechanism(s) involved in the premature destruction of erythrocytes in peripheral blood, implicated in the hemoglobin disorder, HbE/beta-thalassemia. |
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Keywords: | HbA, adult hemoglobin HbF, fetal hemoglobin PC, phosphatidylcholine PE, phosphatidylethanolamine PS, phosphatidylserine Chol, cholesterol DMPC, dimyristoylphosphatidylcholine DMPE, dimyristoylphosphatidylethanolamine DMPS, dimyristoylphosphatidylserine DOPC, dioleoylphosphatidylcholine DOPE, dioleoylphosphatidylethanolamine CF, 6-carboxyfluorescein SUV, small unilamellar vesicles PMB, p-hydroxymercuribenzoic acid sodium salt S.E.M., standard error of the mean TEM, transmission electron microscopy |
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