The D-domain of fibrin : structural and functional studies

 

Show simple item record

dc.contributor.advisor Purvis, Langley R en_ZA
dc.contributor.advisor Berman, Mervyn C en_ZA
dc.contributor.author Purves, Maud en_ZA
dc.date.accessioned 2018-02-01T13:29:57Z
dc.date.available 2018-02-01T13:29:57Z
dc.date.issued 1987 en_ZA
dc.identifier.citation Purves, M. 1987. The D-domain of fibrin : structural and functional studies. University of Cape Town. en_ZA
dc.identifier.uri http://hdl.handle.net/11427/27202
dc.description.abstract The D-domain of fibrin (ogen) was separated from the parent molecule by plasmin digestion in the presence of calcium and isolated by means of DEAE-anion exchange chromatography followed by gel-filtration in buffer containing 4 M urea. Fluorescent-D-dimer (f-D-dimer) was isolated from a plasmic digest of fibrin clotted in the presence of 2.45 mM dansyl cadaverine, a fluorescent lysine analogue. Fluorescent-D-monomer was a by-product of f-D-dimer purification, the yield being determined by the concentration of dansyl cadaverine. At 2.45 mM f-D-monomer was always present in the digest. The f-D-monomer is probably formed directly and not as a result of degradation of f-D-dimer. The molecule elutes in the fibrinogen-derived-D- monomer position on gel-filtration. A protease was isolated and partially purified from venom of the puffadder (Bitis arietans). Puffadder venom protease is characterized by its ability to cleave D-dimer into symmetrical D-monomers, smaller than plasmin-derived D-monomers from fibrinogen. This characteristic was used to detect the puffadder venom protease activity with fluorescent-D-dimer being used as the substrate. Fractions obtained were assayed for D-dimer cleavage activity and the samples analyzed by means of SDS-PAGE on 4-20% gradient gels under reducing (βME) and non-reducing conditions. The fluorescent bands were located under U.V light and photographed prior to staining with Coomassie Blue. Several methods for the purification of the protease were investigated. en_ZA
dc.language.iso eng en_ZA
dc.subject.other Fibrin en_ZA
dc.subject.other Fibrin - analysis en_ZA
dc.title The D-domain of fibrin : structural and functional studies en_ZA
dc.type Doctoral Thesis
uct.type.publication Research en_ZA
uct.type.resource Thesis en_ZA
dc.publisher.institution University of Cape Town
dc.publisher.faculty Faculty of Health Sciences en_ZA
dc.publisher.department Division of Chemical Pathology en_ZA
dc.type.qualificationlevel Doctoral
dc.type.qualificationname PhD en_ZA
uct.type.filetype Text
uct.type.filetype Image
dc.identifier.apacitation Purves, M. (1987). <i>The D-domain of fibrin : structural and functional studies</i>. (Thesis). University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology. Retrieved from http://hdl.handle.net/11427/27202 en_ZA
dc.identifier.chicagocitation Purves, Maud. <i>"The D-domain of fibrin : structural and functional studies."</i> Thesis., University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1987. http://hdl.handle.net/11427/27202 en_ZA
dc.identifier.vancouvercitation Purves M. The D-domain of fibrin : structural and functional studies. [Thesis]. University of Cape Town ,Faculty of Health Sciences ,Division of Chemical Pathology, 1987 [cited yyyy month dd]. Available from: http://hdl.handle.net/11427/27202 en_ZA
dc.identifier.ris TY - Thesis / Dissertation AU - Purves, Maud AB - The D-domain of fibrin (ogen) was separated from the parent molecule by plasmin digestion in the presence of calcium and isolated by means of DEAE-anion exchange chromatography followed by gel-filtration in buffer containing 4 M urea. Fluorescent-D-dimer (f-D-dimer) was isolated from a plasmic digest of fibrin clotted in the presence of 2.45 mM dansyl cadaverine, a fluorescent lysine analogue. Fluorescent-D-monomer was a by-product of f-D-dimer purification, the yield being determined by the concentration of dansyl cadaverine. At 2.45 mM f-D-monomer was always present in the digest. The f-D-monomer is probably formed directly and not as a result of degradation of f-D-dimer. The molecule elutes in the fibrinogen-derived-D- monomer position on gel-filtration. A protease was isolated and partially purified from venom of the puffadder (Bitis arietans). Puffadder venom protease is characterized by its ability to cleave D-dimer into symmetrical D-monomers, smaller than plasmin-derived D-monomers from fibrinogen. This characteristic was used to detect the puffadder venom protease activity with fluorescent-D-dimer being used as the substrate. Fractions obtained were assayed for D-dimer cleavage activity and the samples analyzed by means of SDS-PAGE on 4-20% gradient gels under reducing (βME) and non-reducing conditions. The fluorescent bands were located under U.V light and photographed prior to staining with Coomassie Blue. Several methods for the purification of the protease were investigated. DA - 1987 DB - OpenUCT DP - University of Cape Town LK - https://open.uct.ac.za PB - University of Cape Town PY - 1987 T1 - The D-domain of fibrin : structural and functional studies TI - The D-domain of fibrin : structural and functional studies UR - http://hdl.handle.net/11427/27202 ER - en_ZA


Files in this item

This item appears in the following Collection(s)

Show simple item record