Metals and the conformation of fibrin

Simple item page
dc.contributor.advisorPurvis, Langley Ren_ZA
dc.contributor.authorNaidoo, Dhesigen Pen_ZA
dc.date.accessioned2017-12-12T10:55:32Z
dc.date.available2017-12-12T10:55:32Z
dc.date.issued1992en_ZA
dc.description.abstractThe carboxy terminal of the γ-chain of human fibrinogen contains at least three biologically. important functional domains: (i) the fibrinogen γ-chain polymerisation centre, (ii) the platelet receptor domain and (iii) the site for staphyloccocal clumping. The nature of the site specificity of these interactions necessitates the existence of a preferred conformation for this region, the nature of which has yet to be clearly established. A novel zinc metalloproteinase isolated from puff adder venom (PAV protease) capable of specifically cleaving the di-γ-chain of transglutaminase (Factor XIIIa) catalysed crosslinked plasmin derived D-dimer into apparently symmetrical monomers has been described. The activity is fibrin specific and displays an unusual site specificity for the γ-carboxy terminal domains within the crosslink region. The activity was reported to be potentiated by zinc. The effect of zinc on the digestion of D-dimer by PAV protease was evaluated by SDS-PAGE and by a fluorimetric technique utilising a fluorescent dansylcadaverine conjugate of the substrate (f-D-dimer). A differential zinc binding study determined that the potentiation of activity by zinc was due to a zinc-substrate rather than a zinc-enzyme interaction. The binding constant for zinc to D-dimer was determined by Scatchard analysis of zinc titration data. The interaction of zinc and f-D-dimer was confirmed by fluorescence anisotropy determinations. The nature of the coordination capsule around the metal cation was determined by examining a cobalt-fibrin-D-dimer complex and characterising the difference visible absorption spectrum thereof. The donor ligands from the D-dimer fragment for the metal ion were determined as histidines by examining zinc(II) and cobalt(II) binding to diethylpyrocarbonate modified fibrin-D-dimer and hydroxylamine treated DEPC-fibrin-D-dimer. Through this study it has been established that the PAV protease cleavage of the di-γ-chain of the plasmin derived D-dimer fragment is potentiated by zinc(II) ions through the formation of a novel zinc determined conformation of fibrin-D-dimer. This presents the possibility of a fibrinspecific neo-epitope being manifested in the presence of zinc ions that could provide a means to determine fibrin degradation products more specifically. A model for the neo-epitope has been proposed.en_ZA
dc.identifier.apacitationNaidoo, D. P. (1992). <i>Metals and the conformation of fibrin</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology. Retrieved from http://hdl.handle.net/11427/26555en_ZA
dc.identifier.chicagocitationNaidoo, Dhesigen P. <i>"Metals and the conformation of fibrin."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1992. http://hdl.handle.net/11427/26555en_ZA
dc.identifier.citationNaidoo, D. 1992. Metals and the conformation of fibrin. University of Cape Town.en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Naidoo, Dhesigen P AB - The carboxy terminal of the γ-chain of human fibrinogen contains at least three biologically. important functional domains: (i) the fibrinogen γ-chain polymerisation centre, (ii) the platelet receptor domain and (iii) the site for staphyloccocal clumping. The nature of the site specificity of these interactions necessitates the existence of a preferred conformation for this region, the nature of which has yet to be clearly established. A novel zinc metalloproteinase isolated from puff adder venom (PAV protease) capable of specifically cleaving the di-γ-chain of transglutaminase (Factor XIIIa) catalysed crosslinked plasmin derived D-dimer into apparently symmetrical monomers has been described. The activity is fibrin specific and displays an unusual site specificity for the γ-carboxy terminal domains within the crosslink region. The activity was reported to be potentiated by zinc. The effect of zinc on the digestion of D-dimer by PAV protease was evaluated by SDS-PAGE and by a fluorimetric technique utilising a fluorescent dansylcadaverine conjugate of the substrate (f-D-dimer). A differential zinc binding study determined that the potentiation of activity by zinc was due to a zinc-substrate rather than a zinc-enzyme interaction. The binding constant for zinc to D-dimer was determined by Scatchard analysis of zinc titration data. The interaction of zinc and f-D-dimer was confirmed by fluorescence anisotropy determinations. The nature of the coordination capsule around the metal cation was determined by examining a cobalt-fibrin-D-dimer complex and characterising the difference visible absorption spectrum thereof. The donor ligands from the D-dimer fragment for the metal ion were determined as histidines by examining zinc(II) and cobalt(II) binding to diethylpyrocarbonate modified fibrin-D-dimer and hydroxylamine treated DEPC-fibrin-D-dimer. Through this study it has been established that the PAV protease cleavage of the di-γ-chain of the plasmin derived D-dimer fragment is potentiated by zinc(II) ions through the formation of a novel zinc determined conformation of fibrin-D-dimer. This presents the possibility of a fibrinspecific neo-epitope being manifested in the presence of zinc ions that could provide a means to determine fibrin degradation products more specifically. A model for the neo-epitope has been proposed. DA - 1992 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1992 T1 - Metals and the conformation of fibrin TI - Metals and the conformation of fibrin UR - http://hdl.handle.net/11427/26555 ER - en_ZA
dc.identifier.urihttp://hdl.handle.net/11427/26555
dc.identifier.vancouvercitationNaidoo DP. Metals and the conformation of fibrin. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1992 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/26555en_ZA
dc.language.isoengen_ZA
dc.publisher.departmentDivision of Chemical Pathologyen_ZA
dc.publisher.facultyFaculty of Health Sciencesen_ZA
dc.publisher.institutionUniversity of Cape Town
dc.subject.otherFibrin - chemistryen_ZA
dc.subject.otherMetals - Analysisen_ZA
dc.subject.otherZinc - analysisen_ZA
dc.titleMetals and the conformation of fibrinen_ZA
dc.typeMaster Thesis
dc.type.qualificationlevelMasters
dc.type.qualificationnameMSc (Med)en_ZA
uct.type.filetypeText
uct.type.filetypeImage
uct.type.publicationResearchen_ZA
uct.type.resourceThesisen_ZA
Files
Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
thesis_hsf_1992_naidoo_dhesigen_p (1).pdf
Size:
9.76 MB
Format:
Adobe Portable Document Format
Description:
Download
Collections
Masters