Schistosoma mansoni and Fasciola hepatica are parasitic trematodes which contain cytochrome c peroxidase (CcP) in their mitochondria, an enzyme that is absent in mammalian tissues. CcP reduces hydrogen peroxide to H2O using cytochrome c as the electron donor. Both parasites are catalase deficient; thus, cytochrome c peroxidase and glutathione peroxidase are the enzymes involved in the detoxification of H2O 2 in these organisms. The enzymatic activity of these two peroxidases may enable S. mansoni and F. hepatica to survive oxidative stress. The main objective of this study was to characterize cytochrome c peroxidase from S. mansoni and F. hepatica . Kinetic studies of this enzyme in crude homogenate and isolated mitochondria of S. mansoni were initially performed, followed by purification studies from S. mansoni and F. hepatica . The parasite enzyme has affinity for horse heart and yeast cytochrome c and it is inhibited by sodium azide and potassium cyanide. CcP was purified close to homogeneity and identified as a protein containing heme. The antioxidant capability of F. hepatica CcP was tested in vitro , demonstrating that CcP protected the sugar deoxyribose from oxidative degradation. Exposure of adult worms to H2O2 caused a decrease in S. mansoni CcP activity in vivo. An attempt was made to clone the S. mansoni CcP gene. The experiments did not result in the cloning of the CcP gene, but led to the identification and cloning of another protein, a component of a cytosolic chaperonin, t-complex polypeptide one (TCP-1). TCP-1 from S. mansoni is highly homologous to TCP-1 proteins from different organisms including, Chinese hamster, human, Drosophila and yeast and carries ATP binding amino acid motifs indicating that it has ATPase activity.

Hsp104 and ClpB are hexameric ATPases that resolubilize aggregated proteins in collaboration with the Hsp70 chaperone system. Hsp104/ClpB functionally interact only with their respective Hsp70 system and this specificity is mapped to the Hsp104/ClpB coiled-coil domain (CCD). We hypothesize that the interaction between Hsp70 and Hsp104/ClpB CCD stimulates nucleotide exchange and release of substrate from Hsp70. In the current study, the CCDs of E. coli ClpB and S. cerevisiae Hsp104 have been purified. Isolated domains are monomeric and well folded. They inhibit refolding of aggregated firefly luciferase in a species-specific manner. We found that the ATPase activity of E. coli DnaK is stimulated at low concentrations of the E. coli ClpB CCD but not by yeast Hsp104 CCD. However, in another bacterial system (Thermus thermophilus) we found that the ClpB CCD inhibits The ATPase activity of DnaK suggesting that the interaction may have different consequences in distinct chaperone networks.

Thesis (Ph.D.)--University of Wollongong, 2007. / Typescript. Includes bibliographical references: leaf 220-255.

Thesis (M.Sc.) - Carleton University, 2006. / Includes bibliographical references (p. 151-170). Also available in electronic format on the Internet.

Thesis (Ph.D. in Biochemistry & Molecular Genetics) -- University of Colorado at Denver and Health Sciences Center, 2006. / Typescript. Includes bibliographical references (leaves 169-185). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

Thesis (Ph. D.)--University of Washington, 1998. / Vita. Includes bibliographical references (leaves [167]-182).

Thesis (M.Sc.)--University of Wollongong, 2008. / Typescript. Includes bibliographical references (p. 107-120)

Thesis (M.S.) -- Worcester Polytechnic Institute. / Keywords: metal transport; Cu+; Cys; PIB-type ATPase; CopA. Includes bibliographical references (p. 39-45).

Thesis (Ph. D. (Biochemistry, Microbiology & Biotechnology))--Rhodes University, 2004.

Thesis (Ph.D. in Biochemistry) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 128-144). Free to UCD affiliates. Online version available via ProQuest Digital Dissertations;