Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease

dc.contributor.advisorRyall, Roseen_ZA
dc.contributor.authorWebber, Dawnen_ZA
dc.date.accessioned2016-04-22T13:40:24Z
dc.date.available2016-04-22T13:40:24Z
dc.date.issued2003en_ZA
dc.description.abstractThe incidence of kidney stones amongst South Africa's black population is rare. This is in contrast to the white population, whose stone rate is similar to that in Western society. Urine composition alone does not account for these differences. This thesis presents a study of the inhibitory role of the protein, urinary prothrombin fragment 1 (UPTFI ), and its biochemical characterisation in both population groups. In a preliminary study, the urine composition and inhibitory activity of urine and urinary macromolecules from healthy white and black subjects was compared using a spectrophotometric sedimentation assay, zeta potential measurements and particle size analysis. Results suggested greater inhibition by urinary macromolecules in the black group. UPTFI was isolated from calcium oxalate (CaOx) crystals and purified by reverse phase (RP)-high performance liquid chromatography (HPLC) from the urine of healthy white (WFl) and black (BFl) subjects. The identity of the purified proteins was confirmed by Western blotting, N-terminal protein sequencing, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry (MS), amino acid analysis and 2D sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE); these analyses did not indicate differences in the protein backbone from the two groups. However, alkaline amino acid analysis showed the presence of more y-carboxyglutamic acid (Gla) residues in BFI. The N-and 0-linked glycans were released by enzymatic and chemical reactions, respectively, and sequenced using exoglycosidase digestions in tandem with RP and weak anion exchange HPLC, as well as MS. These analyses demonstrated a high proportion of sialylated glycans on UPTFl and a greater number of sialic acid residues on BFI. Molecular modeling located the glycans on the protein's kringle domain and identified a potential mode by which crystallisation could be inhibited.en_ZA
dc.identifier.apacitationWebber, D. (2003). <i>Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease</i>. (Thesis). University of Cape Town ,Faculty of Science ,Department of Chemistry. Retrieved from http://hdl.handle.net/11427/19148en_ZA
dc.identifier.chicagocitationWebber, Dawn. <i>"Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease."</i> Thesis., University of Cape Town ,Faculty of Science ,Department of Chemistry, 2003. http://hdl.handle.net/11427/19148en_ZA
dc.identifier.citationWebber, D. 2003. Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Webber, Dawn AB - The incidence of kidney stones amongst South Africa's black population is rare. This is in contrast to the white population, whose stone rate is similar to that in Western society. Urine composition alone does not account for these differences. This thesis presents a study of the inhibitory role of the protein, urinary prothrombin fragment 1 (UPTFI ), and its biochemical characterisation in both population groups. In a preliminary study, the urine composition and inhibitory activity of urine and urinary macromolecules from healthy white and black subjects was compared using a spectrophotometric sedimentation assay, zeta potential measurements and particle size analysis. Results suggested greater inhibition by urinary macromolecules in the black group. UPTFI was isolated from calcium oxalate (CaOx) crystals and purified by reverse phase (RP)-high performance liquid chromatography (HPLC) from the urine of healthy white (WFl) and black (BFl) subjects. The identity of the purified proteins was confirmed by Western blotting, N-terminal protein sequencing, matrix-assisted laser desorption ionisation time-of-flight (MALDI-TOF) mass spectrometry (MS), amino acid analysis and 2D sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE); these analyses did not indicate differences in the protein backbone from the two groups. However, alkaline amino acid analysis showed the presence of more y-carboxyglutamic acid (Gla) residues in BFI. The N-and 0-linked glycans were released by enzymatic and chemical reactions, respectively, and sequenced using exoglycosidase digestions in tandem with RP and weak anion exchange HPLC, as well as MS. These analyses demonstrated a high proportion of sialylated glycans on UPTFl and a greater number of sialic acid residues on BFI. Molecular modeling located the glycans on the protein's kringle domain and identified a potential mode by which crystallisation could be inhibited. DA - 2003 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 2003 T1 - Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease TI - Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease UR - http://hdl.handle.net/11427/19148 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/19148
dc.identifier.vancouvercitationWebber D. Urinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone disease. [Thesis]. University of Cape Town ,Faculty of Science ,Department of Chemistry, 2003 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/19148en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDepartment of Chemistryen_ZA
dc.publisher.facultyFaculty of Scienceen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherChemistryen_ZA
dc.titleUrinary prothrombin fragment 1 : a potential role-player in the protection of South African blacks from calcium oxalate kidney stone diseaseen_ZA
dc.typeDoctoral Thesis
dc.type.qualificationlevelDoctoral
dc.type.qualificationnamePhDen_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
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