Equine UDP-glucuronosyltransferases and their role in phase II metabolismTools Pickwell, Natalie Dawn (2018) Equine UDP-glucuronosyltransferases and their role in phase II metabolism. PhD thesis, University of Nottingham.
AbstractMetabolism is an essential chemical process and pathway involving multiple reactions. Oxidative or functionalisation reactions dominate phase 1 metabolism and is predominately controlled by the Cytochrome P450s (P450s). Phase 2 reactions are frequently referred to as the detoxification, or elimination, phase. Several families of enzymes are involved, and the largest of these are the Uridine diphosphate 5’-glucuronosyltransferases (UGTs). The purpose of this project was to initiate the development of an equine in vitro toolbox, concentrating on the UGTs. This required identification of UGTs in the equine genome. The first step used syntenic analysis which enabled us to utilise relative gene order conservation between species to determine whether the predicted gene encoded a member of the UGT superfamily. Further analysis of sequence relationships provided confidence that the genes under investigation were UGTs, but also allowed us to determine which UGT orthologue we were investigating. PCRs were performed to isolate the genes, and subsequent sequencing enabled the UGTs to be investigated for key features, including signal peptides, signature sequences, transmembrane domains, and dilysine repeats, which are characteristic of this family of membrane-bound proteins.
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