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Public Health Relevance

Hemoglobin (Hb) E (?E26K) is the most common worldwide naturally occurring mutant Hb with a mutation at the ?1?1 interface. EE individuals exhibit a mild, chronic anemia while HbE/?-thalassemia individuals show a range of clinical manifestations, including high morbidity, and death, often resulting from cardiac dysfunction. The significant role of HbE in the red blood cell pathophysiology and molecular mechanisms giving rise to the HbE diseases is enigmatic. Since, HbE has been shown to have normal oxygen affinity (Bunn et al., 1972), we have proposed a possible mechanism whereby HbE may have reduced capacity to generate sufficient bioactive nitric oxide (NO) (1) to confer protection against high levels of membrane damaging reactive oxygen species (ROS) arising from the ?-thalassemia and (2) as a secondary NO source for endothelial functioning. In support of this hypothesis we have obtained preliminary data from HbE showing decreased nitrite reductase activity compared to HbA. Our group recently obtained the high resolution deoxy and liganded HbE structures (Protein Data Bank entries 1YVQ, 1YVT, 3DUT) and found that the tertiary conformations within the T and R quaternary structures of HbE are altered relative to HbA. The proposed project which builds on these two findings, seeks to establish the extent and molecular origins of the altered nitrite reactivity. The project will utilize innovative sol-gel encapsulation protocols to trap and characterize the reactivity of the T and R state of HbE with respect to a series of reactions proposed to contribute to the production of bioactive NO. The project seeks to determine whether the source of HbE altered reactivity to generate bioactive forms of NO from nitrite arises from changes in allostery, in local tertiary structure or in the redox properties of the T and R states. These studies are of significance in that they are likely to provide a novel mechanism for the origin of HbE-derived pathophysiology with implications for other Hb related pathologies and diseases linked to endothelial dysfunction. There is the potential for a new paradigm in which to develop therapies for HbE ?-thalassemia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HL106421-02
Application #
8206640
Study Section
Molecular and Cellular Hematology (MCH)
Program Officer
Goldsmith, Jonathan C
Project Start
2010-12-15
Project End
2013-07-31
Budget Start
2011-12-01
Budget End
2013-07-31
Support Year
2
Fiscal Year
2012
Total Cost
$207,500
Indirect Cost
$82,500
Name
Albert Einstein College of Medicine
Department
Physiology
Type
Schools of Medicine
DUNS #
110521739
City
Bronx
State
NY
Country
United States
Zip Code
10461
Roche, Camille J; Cassera, Maria B; Dantsker, David et al. (2013) Generating S-nitrosothiols from hemoglobin: mechanisms, conformational dependence, and physiological relevance. J Biol Chem 288:22408-25
Chen, Qiuying; Fabry, Mary E; Rybicki, Anne C et al. (2012) A transgenic mouse model expressing exclusively human hemoglobin E: indications of a mild oxidative stress. Blood Cells Mol Dis 48:91-101
Roche, Camille J; Malashkevich, Vladimir; Balazs, Tatiana C et al. (2011) Structural and functional studies indicating altered redox properties of hemoglobin E: implications for production of bioactive nitric oxide. J Biol Chem 286:23452-66