{"id":{"repo_id":"vt","oai_identifier":"oai:vtechworks.lib.vt.edu:10919/101421"},"canonical_url":"https://search.dev.ndltd.org/etd/vt/oai:vtechworks.lib.vt.edu:10919/101421","repository":{"repo_id":"vt","name":"Virginia Tech","base_url":"https://vtechworks.lib.vt.edu/oai/request"},"display":{"title":"Purification of uridine diphosphate glucuronyltransferase","abstract":"Detoxification of compounds occurs in two phases. In phase I, a functional group on the toxicant is made available for subsequent Phase II reactions. In phase II, the functional groups are conjugated to a compound that will increase the solubility of the toxicant, enhancing its elimination. Uridine diphosphate glucuronyl transferase (UDPGT) is one microsomal enzyme involved in phase II reactions. It catalyzes the conjugation of toxic compounds with glucuronic acid in reactions in which uridine diphosphate glucuronic acid (UDPGA) is the donor substrate. A new purification procedure for UDPGT has been developed. This procedure includes a Polyethylene glycol fractionation, ion exchange chromatography with DEAE Bio-gel A and affinity chromatography with UDP-hexanolamine-Sepharose. The purification was monitored for three different substrates, bilirubin, 4-nitrophenol (PNP) and 7-hydroxy-4-methylcoumarin (HMC). For this last substrate, HMC, a new continuous fluorometric assay was developed. The purification fold and activity recovery, respectively, towards each substrate was as follows: bilirubin, 31 and 80%; PNP, 31 and 80%; and HMC, 24 and 60%. The significance of these results is discussed with reference to the activation of UDPGT in microsomes by detergents and the reactivation of purified UDPGT by phospholipids.","abstract_html":"Detoxification of compounds occurs in two phases. In phase I, a functional group on the toxicant is made available for subsequent Phase II reactions. In phase II, the functional groups are conjugated to a compound that will increase the solubility of the toxicant, enhancing its elimination. Uridine diphosphate glucuronyl transferase (UDPGT) is one microsomal enzyme involved in phase II reactions. It catalyzes the conjugation of toxic compounds with glucuronic acid in reactions in which uridine diphosphate glucuronic acid (UDPGA) is the donor substrate. A new purification procedure for UDPGT has been developed. This procedure includes a Polyethylene glycol fractionation, ion exchange chromatography with DEAE Bio-gel A and affinity chromatography with UDP-hexanolamine-Sepharose. The purification was monitored for three different substrates, bilirubin, 4-nitrophenol (PNP) and 7-hydroxy-4-methylcoumarin (HMC). For this last substrate, HMC, a new continuous fluorometric assay was developed. The purification fold and activity recovery, respectively, towards each substrate was as follows: bilirubin, 31 and 80%; PNP, 31 and 80%; and HMC, 24 and 60%. The significance of these results is discussed with reference to the activation of UDPGT in microsomes by detergents and the reactivation of purified UDPGT by phospholipids.","abstract_has_math":false,"creators":["Duffaud, Guy Daniel"],"institution":"Virginia Polytechnic Institute and State University","degree_name":"M.S.","degree_level":"masters","degree_discipline":"Biochemistry and Nutrition","degree_department":"Biochemistry and Nutrition","school":null,"contributors":[],"advisors":[],"committee_chairs":[],"committee_members":[],"year":1983,"date_issued":"1983","date_published":"1983","updated_at":"2026-07-22T22:19:01Z","subjects":[],"languages":["en"],"rights":["In Copyright"],"rights_urls":["http://rightsstatements.org/vocab/InC/1.0/"],"identifier_entries":[]},"links":{"outbound_url":"http://hdl.handle.net/10919/101421","outbound_label":"Handle","outbound_source":"dc:identifier.uri"},"metadata_groups":[{"id":"people","label":"People","entries":[{"key":"dc:contributor.department","label":"Department","values":["Biochemistry and Nutrition"]},{"key":"dc:creator","label":"Author","values":["Duffaud, Guy Daniel"]}]},{"id":"academic_context","label":"Academic Context","entries":[{"key":"dc:date.accessioned","label":"Dc Date Accessioned","values":["2020-12-15T19:11:31Z"]},{"key":"dc:date.available","label":"Dc Date Available","values":["2020-12-15T19:11:31Z"]},{"key":"dc:date.issued","label":"Date","values":["1983"]},{"key":"dc:publisher","label":"Institution","values":["Virginia Polytechnic Institute and State University"]},{"key":"dc:type","label":"Dc Type","values":["Thesis"]},{"key":"dc:type.dcmitype","label":"Dc Type Dcmitype","values":["Text"]},{"key":"thesis:degree_discipline","label":"Discipline","values":["Biochemistry and Nutrition"]},{"key":"thesis:degree_level","label":"Degree Level","values":["masters"]},{"key":"thesis:degree_name","label":"Degree Name","values":["M.S."]},{"key":"thesis:institution_name","label":"Thesis Institution Name","values":["Virginia Polytechnic Institute and State University"]}]},{"id":"language_rights","label":"Language and Rights","entries":[{"key":"dc:language.iso","label":"Language (ISO)","values":["en"]},{"key":"dc:rights","label":"Dc Rights","values":["In Copyright"]},{"key":"dc:rights.uri","label":"Rights URI","values":["http://rightsstatements.org/vocab/InC/1.0/"]}]},{"id":"identifiers","label":"Identifiers","entries":[{"key":"dc:identifier.uri","label":"Identifier URI","values":["http://hdl.handle.net/10919/101421"]}]},{"id":"additional","label":"Additional Metadata","entries":[{"key":"dc:description.abstract","label":"Abstract","values":["Detoxification of compounds occurs in two phases. In phase I, a functional group on the toxicant is made available for subsequent Phase II reactions. In phase II, the functional groups are conjugated to a compound that will increase the solubility of the toxicant, enhancing its elimination. Uridine diphosphate glucuronyl transferase (UDPGT) is one microsomal enzyme involved in phase II reactions. It catalyzes the conjugation of toxic compounds with glucuronic acid in reactions in which uridine diphosphate glucuronic acid (UDPGA) is the donor substrate. A new purification procedure for UDPGT has been developed. This procedure includes a Polyethylene glycol fractionation, ion exchange chromatography with DEAE Bio-gel A and affinity chromatography with UDP-hexanolamine-Sepharose. The purification was monitored for three different substrates, bilirubin, 4-nitrophenol (PNP) and 7-hydroxy-4-methylcoumarin (HMC). For this last substrate, HMC, a new continuous fluorometric assay was developed. The purification fold and activity recovery, respectively, towards each substrate was as follows: bilirubin, 31 and 80%; PNP, 31 and 80%; and HMC, 24 and 60%. The significance of these results is discussed with reference to the activation of UDPGT in microsomes by detergents and the reactivation of purified UDPGT by phospholipids."]},{"key":"dc:description.degree","label":"Dc Description Degree","values":["M.S."]},{"key":"dc:format.mimetype","label":"Dc Format Mimetype","values":["application/pdf"]},{"key":"dc:title","label":"Title","values":["Purification of uridine diphosphate glucuronyltransferase"]}]}],"canonical_facts":{"dc:contributor.department":["Biochemistry and Nutrition"],"dc:creator":["Duffaud, Guy Daniel"],"dc:date.accessioned":["2020-12-15T19:11:31Z"],"dc:date.available":["2020-12-15T19:11:31Z"],"dc:date.issued":["1983"],"dc:description.abstract":["Detoxification of compounds occurs in two phases. In phase I, a functional group on the toxicant is made available for subsequent Phase II reactions. In phase II, the functional groups are conjugated to a compound that will increase the solubility of the toxicant, enhancing its elimination. Uridine diphosphate glucuronyl transferase (UDPGT) is one microsomal enzyme involved in phase II reactions. It catalyzes the conjugation of toxic compounds with glucuronic acid in reactions in which uridine diphosphate glucuronic acid (UDPGA) is the donor substrate. A new purification procedure for UDPGT has been developed. This procedure includes a Polyethylene glycol fractionation, ion exchange chromatography with DEAE Bio-gel A and affinity chromatography with UDP-hexanolamine-Sepharose. The purification was monitored for three different substrates, bilirubin, 4-nitrophenol (PNP) and 7-hydroxy-4-methylcoumarin (HMC). For this last substrate, HMC, a new continuous fluorometric assay was developed. The purification fold and activity recovery, respectively, towards each substrate was as follows: bilirubin, 31 and 80%; PNP, 31 and 80%; and HMC, 24 and 60%. The significance of these results is discussed with reference to the activation of UDPGT in microsomes by detergents and the reactivation of purified UDPGT by phospholipids."],"dc:description.degree":["M.S."],"dc:format.mimetype":["application/pdf"],"dc:identifier.uri":["http://hdl.handle.net/10919/101421"],"dc:language.iso":["en"],"dc:publisher":["Virginia Polytechnic Institute and State University"],"dc:rights":["In Copyright"],"dc:rights.uri":["http://rightsstatements.org/vocab/InC/1.0/"],"dc:title":["Purification of uridine diphosphate glucuronyltransferase"],"dc:type":["Thesis"],"dc:type.dcmitype":["Text"],"thesis:degree_discipline":["Biochemistry and Nutrition"],"thesis:degree_level":["masters"],"thesis:degree_name":["M.S."],"thesis:institution_name":["Virginia Polytechnic Institute and State University"]},"updated_at":"2026-07-22T22:19:01Z"}