The pathological manifestations of sickle cell anemia arises from the polymerization of HbS into long fibers which distort the shape of the red blood cell. The resulting abnormal shape and increased rigidity of the red blood cells impedes blood flow through the capillaries causing the vasoocclusive complications that characterize sickle cell anemia. Cerebral vascular disease is a common and one of many catastrophic complications in children suffering from sickle cell anemia. We have started a long-term investigation into the structure and functional of enzymes in the prostaglandin synthesis cascade. Most of these enzymes are membrane bound enzymes and thus require special handing to prepare them for biophysical analysis. As we have developed methods for crystallizing membrane proteins, they now can be made amendable to X-ray diffraction analysis. Using X- ray crystallography, electron paramagnetic resonance, UV-Vis absorption spectroscopy and fluorine-NMR, we intend to study the enzymes that participate in the synthesis of vasoregulatory prostaglandins. We are now investigating enzyme-drug and enzyme- substrate analog complexes of prostagland H synthase (cyclo- oxygenase), both in solution and in crystals. The products of prostaglandin H synthase, PGH1 and PGH2, serve as precursors for synthesis of prostaglandins which affect platelet aggression and vasodilation. We intend to use the methods we have developed for the study prostaglandin synthase to purify, crystalize and investigate and investigate PGH-PGE isomerase and PGI2 synthase which synthesize powerful vasodilators and inhibitors of platelet aggregation. Our long term goal is to elucidate the structural mechanisms of substrate, product and inhibitor binding to 1) understand better the molecular events occurring during catalysis and 2) to provide the necessary structural information for the design of drugs to control the synthesis of specific prostaglandin. Having inhibitors or activators of the synthesis of specific prostaglandins could allow the development of rational drug therapy for vasoocclusive diseases like sickle cell anemia.

Agency
National Institute of Health (NIH)
Institute
National Heart, Lung, and Blood Institute (NHLBI)
Type
Research Program Projects (P01)
Project #
5P01HL030121-09
Application #
3844258
Study Section
Project Start
Project End
Budget Start
Budget End
Support Year
9
Fiscal Year
1992
Total Cost
Indirect Cost
Name
University of Chicago
Department
Type
DUNS #
225410919
City
Chicago
State
IL
Country
United States
Zip Code
60637
Cassady, Kevin A; Gross, Martin (2002) The herpes simplex virus type 1 U(S)11 protein interacts with protein kinase R in infected cells and requires a 30-amino-acid sequence adjacent to a kinase substrate domain. J Virol 76:2029-35
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Cassady, K A; Gross, M; Roizman, B (1998) The second-site mutation in the herpes simplex virus recombinants lacking the gamma134.5 genes precludes shutoff of protein synthesis by blocking the phosphorylation of eIF-2alpha. J Virol 72:7005-11
Cassady, K A; Gross, M; Roizman, B (1998) The herpes simplex virus US11 protein effectively compensates for the gamma1(34.5) gene if present before activation of protein kinase R by precluding its phosphorylation and that of the alpha subunit of eukaryotic translation initiation factor 2. J Virol 72:8620-6
He, B; Gross, M; Roizman, B (1998) The gamma134.5 protein of herpes simplex virus 1 has the structural and functional attributes of a protein phosphatase 1 regulatory subunit and is present in a high molecular weight complex with the enzyme in infected cells. J Biol Chem 273:20737-43
He, B; Gross, M; Roizman, B (1997) The gamma(1)34.5 protein of herpes simplex virus 1 complexes with protein phosphatase 1alpha to dephosphorylate the alpha subunit of the eukaryotic translation initiation factor 2 and preclude the shutoff of protein synthesis by double-stranded RNA-activa Proc Natl Acad Sci U S A 94:843-8
Picot, D; Loll, P J; Garavito, R M (1997) X-ray crystallographic study of the structure of prostaglandin H synthase. Adv Exp Med Biol 400A:107-11
Goldwasser, E (1996) Erythropoietin: a somewhat personal history. Perspect Biol Med 40:18-32

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