The isolation and charcterisation of ovine liver cytochrome b₅

Lombard, Nicolaas (2007-03)

Dissertation (PhD)--University of Stellenbosch, 2007.


ENGLISH ABSTRACT: This dissertation describes how the isolation and characterisation of ovine liver cytochrome b5 was accomplished by referring to the following goals achieved in this study: - The optimisation of the isolation and purification procedure for ovine liver microsomal cytochrome b5 in order to obtain sufficient material for aggregation and immunological studies. - The removal of the membrane binding domain of cytochrome b5 by means of tryptic digestion to establish the role of the carboxyl terminal in ovine cytochrome b5 aggregation. - The raising of antibodies against both the trypsin truncated and intact forms of cytochrome b5 to study the aggregation of the protein. - The investigation into the influence of purified cytochrome b5 on steroidogenesis in ovine adrenal microsomes.

AFRIKAANSE OPSOMMING: Die isolering en karakterisering van skaaplewersitochroom b5, soos beskryf in hierdie proefskrif, is uitgevoer deur die volgende doelwitte suksesvol af te handel: - Die optimalisering van die prosedure vir die suksesvolle isolering en suiwering van skaaplewersitochroom b5 ten einde genoegsame hoeveelhede van die suiwer proteïen te hê vir die bestudering van die aggregasie van die proteïen sowel as ‘n immunologiese studie. - Die verwydering van die membraanbindingsdomein van sitochroom b5 om die invloed van die karboksielterminaal op die aggregering van die proteïen te bestudeer. - Die gebruik van sowel die tripties gesnyde as die intakte vorms van sitochroom b5 om ‘n immuunrespons in hase op te wek vir die verkryging van sitochroom b5 spesifieke anti-liggame. - Die gebruik van die gesuiwerde proteïene om die invloed van sitochroom b5 op adrenale steroïdogenese te bestudeer.

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