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Spin Label Studies of the Hemoglobin–Membrane Interaction During Sickle Hemoglobin Polymerization

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Abstract

An enhanced hemoglobin–membrane association has been previously documented in sickle cell anemia. However, it is not known how this interaction is modified during the hemoglobin S polymerization process. In this work, we use a model of reconstituted erythrocytes from ghost membranes whose cytoskeleton proteins had been previously labeled with the 4-maleimido Tempo spin label, and that were subsequently resealed with hemoglobin S or A solutions. Using electron paramagnetic resonance spectroscopy, we studied the time dependence of the spectral W/S parameter, indicative of the conformational state of cytoskeleton proteins (mainly spectrin) under spontaneous deoxygenation, with the aim of detecting the eventual effects due to hemoglobin S polymerization. The differences observed in the temporal behavior of W/S in erythrocytes reconstituted with both hemoglobins were considered as experimental evidence of an increment in hemoglobin S–membrane interaction as a result of the polymerization process of hemoglobin S under spontaneous deoxygenation.

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Acknowledgments

Financial support for this work was provided by the Binational Cooperation Program MINCYT (Argentina)-CITMA (Cuba), Universidad de Oriente (Santiago de Cuba, Cuba), Universidad Nacional del Litoral (Santa Fe, Argentina) and CONICET (Argentina). A.M.G. is a researcher and P.M.R. holds a scholarship from CONICET. We thank biochemist A. Sartore for supplying the blood samples of SCA patients. Thanks also are due to the Abdus Salam International Centre for Theoretical Physics (ICTP), Trieste, especially to the Associateship Scheme, in which J.E.F.D. is a junior associate member.

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Authors and Affiliations

  1. Centro de Biofísica Médica, Universidad de Oriente, Patricio Lumumba s/n, 90500, Santiago de Cuba, Cuba

    José Ernesto Falcón Dieguez & Manuel A. Lores Guevara

  2. INTEC (CONICET-UNL), Guemes 3450, 3000, Santa Fe, Argentina

    Pablo Rodi & Ana Maria Gennaro

  3. Departamento de Física, Facultad de Bioquímica y Ciencias Biológicas, Universidad Nacional del Litoral (UNL), Ciudad Universitaria, 3000, Santa Fe, Argentina

    Pablo Rodi & Ana Maria Gennaro

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  1. José Ernesto Falcón Dieguez
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  2. Pablo Rodi
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  3. Manuel A. Lores Guevara
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  4. Ana Maria Gennaro
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Correspondence to José Ernesto Falcón Dieguez.

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Falcón Dieguez, J.E., Rodi, P., Lores Guevara, M.A. et al. Spin Label Studies of the Hemoglobin–Membrane Interaction During Sickle Hemoglobin Polymerization. Appl Magn Reson 38, 443–453 (2010). https://doi.org/10.1007/s00723-010-0138-8

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  • DOI: https://doi.org/10.1007/s00723-010-0138-8

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