In this project, Dr. Josephs proposes to extend his extensive and elegant earlier work, that has culminated in a highly detailed model for the structure of the sickle cell hemoglobin fiber. This model provides the foundation for understanding the interactions that drive fiber formation and, thus, the clinical manifestations of sickle-cell anemia. The fundamental unit of the sickle-cell fiber is a double-strand of hemoglobin molecules that has been observed in crystals of deoxy hemoglobin molecules S (HbS). The fiber is composed of seven of these double strands that have a helical twist in the fiber, but not in the crystal. The crystal structure analysis by Love and colleagues delineates the interactions within the double strand, however the interactions between double strands that stabilize the fiber are not present in the crystal lattice. Thus, EM analysis, in conjunction with model building, is required to determine the """"""""inter double strand"""""""" contacts. Two competing models have been proposed- one by the group of Dr. Josephs and the other by Edelstein and colleagues. With the use of an efficient microbial systems for production of recombinant hemoglobin, these alternate models can now be tested by site- directed mutagenesis. The proposed project contains three specific aims.
| Yosmanovich, Donna; Rotter, Maria; Aprelev, Alexey et al. (2016) Calibrating Sickle Cell Disease. J Mol Biol 428:1506-14 |
| Tokarev, Alexander; Aprelev, Alexey; Zakharov, Mikhail N et al. (2012) Multifunctional magnetic rotator for micro and nanorheological studies. Rev Sci Instrum 83:065110 |
| Manning, James M; Popowicz, Anthony M; Padovan, Julio C et al. (2012) Intrinsic regulation of hemoglobin expression by variable subunit interface strengths. FEBS J 279:361-9 |
| Weng, Weijun; Ferrone, Frank A (2011) Metastable gels: A novel application of Ogston theory to sickle hemoglobin polymers. Biophys Chem 154:99-101 |
| Rotter, Maria; Yosmanovich, Donna; Briehl, Robin W et al. (2011) Nucleation of sickle hemoglobin mixed with hemoglobin A: experimental and theoretical studies of hybrid-forming mixtures. Biophys J 101:2790-7 |
| Rotter, Maria A; Chu, Haiyan; Low, Philip S et al. (2010) Band 3 catalyzes sickle hemoglobin polymerization. Biophys Chem 146:55-9 |
| Zakharov, Mikhail N; Aprelev, Alexey; Turner, Matthew S et al. (2010) The microrheology of sickle hemoglobin gels. Biophys J 99:1149-56 |
| Manning, Lois R; Popowicz, Anthony M; Padovan, Julio et al. (2010) Developmental expression of human hemoglobins mediated by maturation of their subunit interfaces. Protein Sci 19:1595-9 |
| Wang, Jiang Cheng; Kwong, Suzanna; Ferrone, Frank A et al. (2009) Fiber depolymerization: fracture, fragments, vanishing times, and stochastics in sickle hemoglobin. Biophys J 96:655-70 |
| Manning, Lois R; Russell, J Eric; Popowicz, Anthony M et al. (2009) Energetic differences at the subunit interfaces of normal human hemoglobins correlate with their developmental profile. Biochemistry 48:7568-74 |
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