Thesis (Ph.D.)--McMaster University, 2003. / Advisor: V.S. Ananthanarayanan. Includes bibliographical references. Also available via World Wide Web.

Thesis (M.S.)--University of North Carolina Wilmington, 2007. / Includes bibliographical references (leaves: 190-193)

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 139-147).

Thesis (Ph. D.)--University of Delaware, 2006. / Principal faculty advisor: Brian J. Bahnson, Dept. of Chemistry & Biochemistry. Includes bibliographical references.

No description available.

Vitamin D-binding protein (DBP) is an abundant serum protein, secreted by the liver, which transports vitamin D sterols and is part of an actin scavenging system. In this study, DBP was isolated from horse plasma in a highly reproducible, four step procedure: Affi-gel Blue affinity chromatography, gel filtration, hydroxy1apatite chromatography and anion exchange HPLC. 6-7 mg of DBP were obtained from 80 ml of plasma with a yield of 21-25%. The secondary structure of DBP was calculated from circular dichroism measurements to be 39% α-helix, 42% β-sheet and 19% random coil. A molecular mass of 53,000 ± 3,000 daltons was calculated from electrophoretic gels. Circular dichroism and fluorescence studies revealed that the disulphide bonds of DBP contribute substantial structural stabilization to the molecule with respect to thermal denaturation. Finally, acrylodan-labeled DBP was prepared. The fluorescence of this adduct was sensitive to the binding of actin and to the presence of dithiothreitol. / Science, Faculty of / Chemistry, Department of / Graduate

G proteins are alpha-beta-gamma heterotrimers in the resting state, bound to GDP and complexed with the unbound receptor. Once the receptor becomes occupied, the alpha subunit exchanges GDP for GTP, becomes activated, and dissociates from the receptor and can stimulate or inhibit many intracellular activities such as phosphorylation and channel conductance. For example, Gs and Golf alpha subunits stimulate and Gi alpha subunits inhibit adenylyl cyclase. Go alpha subunits are abundant in brain, but are of unknown function. cDNAs for the alpha subunit have been cloned. In order to examine the relative distributions of G proteins in the brain, we used in situ hybridization with radiolabelled synthetic oligonucleotide probes. By using a tyrosine hydroxylase antibody, we found that the dopaminergic neurons of the substantia nigra and the noradrenergic neurons of the locus ceruleus express mRNA for the alpha subunits for each of Gi, Go, and Gs. We noted a paucity of Gs mRNA in the striatum. This was surprising because the basal ganglia contain a dopamine-stimulated adenylyl cyclase activity which has been assumed to be transduced by Gs. Also, immunohistochemistry, immunoblotting, and cholera ADP-ribosylation indicated a very high level of Gs alpha-like protein in the striatum. In order to ascertain which specific G protein we were detecting, we made probes to a new G protein previously identified in the olfactory system. Golf is a stimulatory G protein with size and sequence characteristics similar to those of Gs. The cholera toxin ADP-ribosylation site and C-terminal region to which the antibody was made are identical. We made oligonucloetide probes to the translated and untranslated portions of Golf alpha. High levels Golf mRNA and protein were detected in the striatum and nucleus accumbens, in addition to the expected high levels in the olfactory tubercle. Northern blot studies indicated that Golf transcripts are approximately ten-fold more abundant than Gs alpha transcripts in the striatum. These data indicate that Golf in not an olfactory-specific G protein. It is also the major stimulatory G protein in the basal ganglia. The selective expression of high levels of Golf in dopamine-rich forebrain areas suggest that it may couple DI dopamine receptors to adenylyl cyclase. The role of Golf in dopaminergic neurotransmission and neuropsychiatric disease should be considered. / Medicine, Faculty of / Graduate

No description available.

No description available.

Azotobacter vinelandii 12837 was grown in Burk's glucose media and transferred onto Burk's n-butanol agar plates to allow for the formation of cysts. The patterns of the vegetative cell proteins were compared for each successive day of cyst formation, using the polyacrylamide gel isoelectric focusing technique. The findings revealed that, as the cysts developed to maturity, definite changes occurred in the protein constitution, indicative of the biochemical and physiological changes which cells undergo during cyst development. Also, as a control to show that the changes in protein patterns during encystment were not due to physiological condition, Azotobacter vinelandii strain OP was grown in three different media, and proteins from the cells were compared using PAGIF.