Band 3 is the major protein of the human erythrocyte membrane and immunologically cross-reactive species have been identified also in nonerythroid cells. The long term goal of this laboratory is to more thoroughly characterize the structure and function of band 3 in red blood cells and to establish the existence of homologous proteins with a similar structure and function in nonerythroid cells. The three major objectives of the current proposal are: i) to crystallize the isolated cytoplasmic domain of band 3 and solve its crystal structure, ii) to locate the subsites on the cytoplasmic domain of band 3 which bind the major cytoskeletal anchoring proteins, ankyrin and band 4.1, and then to investigate the regulation of these interactions, and iii) to screen nonerythroid cells for band 3-like proteins and then to isolate and characterize one or more of these homologous polypeptides. Because the function and/or malfunction of band 3 is reported to be involved in i) senescent red cell removal, ii) several types of hereditary spherocytosis, iii) invasion of the malarial parasite into erythrocytes, iv) attachment of Heinz bodies to the red cell membrane and the consequent hemolytic anemia, v) CO2 transport from the tissues to the lungs as HCO3-, and vi) attachment of the cytoskeleton to the membrane, a more thorough understanding of this protein's structure and function should be relevant to several health fields. The experimental protocols for each of the three main goals listed above involve biochemical techniques which are quite standard for the areas studied. These techniques include: protein crystallization, x-ray diffraction, affinity chromatography, immunoblotting, immunofluorescence microscopy, proteolytic dissection, and simple protein binding assays.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM024417-09
Application #
3272274
Study Section
Cellular Biology and Physiology Subcommittee 1 (CBY)
Project Start
1977-07-01
Project End
1990-03-31
Budget Start
1988-04-01
Budget End
1989-03-31
Support Year
9
Fiscal Year
1988
Total Cost
Indirect Cost
Name
Purdue University
Department
Type
Schools of Arts and Sciences
DUNS #
072051394
City
West Lafayette
State
IN
Country
United States
Zip Code
47907
Giger, Katie; Habib, Ibrahim; Ritchie, Ken et al. (2016) Diffusion of glycophorin A in human erythrocytes. Biochim Biophys Acta 1858:2839-2845
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
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
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
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
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
Sega, Martiana F; Chu, Haiyan; Christian, John et al. (2012) Interaction of deoxyhemoglobin with the cytoplasmic domain of murine erythrocyte band 3. Biochemistry 51:3264-72

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