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
Chu, Haiyan; McKenna, Mary M; Krump, Nathan A et al. (2016) Reversible binding of hemoglobin to band 3 constitutes the molecular switch that mediates O2 regulation of erythrocyte properties. Blood 128:2708-2716
Franco, Taina; Chu, Haiyan; Low, Philip S (2016) Identification of adducin-binding residues on the cytoplasmic domain of erythrocyte membrane protein, band 3. Biochem J 473:3147-58
Puchulu-Campanella, Estela; Turrini, Francesco M; Li, Yen-Hsing et al. (2016) Global transformation of erythrocyte properties via engagement of an SH2-like sequence in band 3. Proc Natl Acad Sci U S A 113:13732-13737
Giger, Katie; Habib, Ibrahim; Ritchie, Ken et al. (2016) Diffusion of glycophorin A in human erythrocytes. Biochim Biophys Acta 1858:2839-2845
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
Sega, Martiana F; Chu, Haiyan; Christian, John A et al. (2015) Fluorescence assay of the interaction between hemoglobin and the cytoplasmic domain of erythrocyte membrane band 3. Blood Cells Mol Dis 55:266-71
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
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
Fernandez-Pol, Sebastian; Slouka, Zdenek; Bhattacharjee, Souvik et al. (2013) A bacterial phosphatase-like enzyme of the malaria parasite Plasmodium falciparum possesses tyrosine phosphatase activity and is implicated in the regulation of band 3 dynamics during parasite invasion. Eukaryot Cell 12:1179-91

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