The structure of the human erythrocyte membrane is described in virtually every modern Cell Biology and Hematology text, because: i) it constitutes a useful model of other plasma membranes, ii) its protein components (or homologues) are present in nearly every cell of the body, iii) its architecture is simple and well characterized, ad iv) defects or alterations in its components lead to important human diseases. The Low lab has focused for 35 years on determining the detailed structure of the red blood cell (RBC) membrane and the impact of defects in its structure on RBC properties. During the course of these studies, the lab has been able to demonstrate that most protein interactions in the membrane are regulated and have significant consequences on RBC properties. However, because the biology of RBC signal transduction has received little attention to date, and since dysfunctions in these signaling pathways can lead to serious human diseases, the goals of this proposal are to characterize two of the most important RBC signaling pathways (i.e. those mediated by O2 and tyrosine phosphorylation) and determine the mechanism by which defects in these pathways contribute to human pathologies.
In Aim 1 transgenic mice will be used to examine whether the oxygen dependent interaction of hemoglobin with band 3 constitutes the "molecular switch" by which oxygen regulates many critical RBC properties.
In Aim 2, the mechanisms by which tyrosine phosphorylation of band 3 induce RBC membrane destabilization and vesiculation will be explored by evaluating protein interactions associated with a unique SH2 domain in band 3.
In Aim 3, a potent inhibitor of this latter pathway will be examined for its impact on the maturation of Plasmodium falciparum within infected red cells and thereby tested as a possible treatment for malaria.

Public Health Relevance

The studies proposed here will investigate the mechanisms by which oxygen pressure and tyrosine phosphorylation regulate red blood cell membrane properties, including membrane stability, membrane vesiculation, cell volume, glucose metabolism, and release of vasodilating agents. Because malfunction of any of these signaling pathways can have serious health consequences, information on the pathways may guide development of new therapies for important human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
2R01GM024417-34
Application #
8575243
Study Section
Molecular and Cellular Hematology (MCH)
Program Officer
Chin, Jean
Project Start
1977-07-01
Project End
2017-05-31
Budget Start
2013-09-30
Budget End
2014-05-31
Support Year
34
Fiscal Year
2013
Total Cost
$444,204
Indirect Cost
$149,554
Name
Purdue University
Department
Chemistry
Type
Schools of Arts and Sciences
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Wandersee, Nancy J; Maciaszek, Jamie L; Giger, Katie M et al. (2015) Dietary supplementation with docosahexanoic acid (DHA) increases red blood cell membrane flexibility in mice with sickle cell disease. Blood Cells Mol Dis 54:183-8
Puchulu-Campanella, Estela; Chu, Haiyan; Anstee, David J et al. (2013) Identification of the components of a glycolytic enzyme metabolon on the human red blood cell membrane. J Biol Chem 288:848-58
Franco, Robert S; Puchulu-Campanella, M Estela; Barber, Latorya A et al. (2013) Changes in the properties of normal human red blood cells during in vivo aging. Am J Hematol 88:44-51
Stefanovic, Marko; Puchulu-Campanella, Estela; Kodippili, Gayani et al. (2013) Oxygen regulates the band 3-ankyrin bridge in the human erythrocyte membrane. Biochem J 449:143-50
Kodippili, Gayani C; Spector, Jeff; Hale, Jacob et al. (2012) Analysis of the mobilities of band 3 populations associated with ankyrin protein and junctional complexes in intact murine erythrocytes. J Biol Chem 287:4129-38
Pantaleo, Antonella; Ferru, Emanuela; Vono, Rosa et al. (2012) New antimalarial indolone-N-oxides, generating radical species, destabilize the host cell membrane at early stages of Plasmodium falciparum growth: role of band 3 tyrosine phosphorylation. Free Radic Biol Med 52:527-36
Ferru, Emanuela; Giger, Katie; Pantaleo, Antonella et al. (2011) Regulation of membrane-cytoskeletal interactions by tyrosine phosphorylation of erythrocyte band 3. Blood 117:5998-6006
Bao, Ning; Kodippili, Gayani C; Giger, Katie M et al. (2011) Single-cell electrical lysis of erythrocytes detects deficiencies in the cytoskeletal protein network. Lab Chip 11:3053-6
Matte, Alessandro; Low, Philip S; Turrini, Franco et al. (2010) Peroxiredoxin-2 expression is increased in beta-thalassemic mouse red cells but is displaced from the membrane as a marker of oxidative stress. Free Radic Biol Med 49:457-66
Kodippili, Gayani C; Spector, Jeff; Kang, Grace E et al. (2010) Analysis of the kinetics of band 3 diffusion in human erythroblasts during assembly of the erythrocyte membrane skeleton. Br J Haematol 150:592-600

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