Your browser doesn't support javascript.
loading
Drug and Chemical Glucosidation by Control Supersomes and Membranes from Spodoptera frugiperda (Sf) 9 Cells: Implications for the Apparent Glucuronidation of Xenobiotics by UDP-glucuronosyltransferase 1A5.
Chau, Nuy; Kaya, Leyla; Lewis, Benjamin C; Mackenzie, Peter I; Miners, John O.
Affiliation
  • Chau N; Department of Clinical Pharmacology (N.C., L.K., B.C.L., P.I.M., J.O.M.) and Flinders Centre for Innovation in Cancer (B.C.L., P.I.M., J.O.M.), Flinders University College of Medicine and Public Health, Adelaide, Australia.
  • Kaya L; Department of Clinical Pharmacology (N.C., L.K., B.C.L., P.I.M., J.O.M.) and Flinders Centre for Innovation in Cancer (B.C.L., P.I.M., J.O.M.), Flinders University College of Medicine and Public Health, Adelaide, Australia.
  • Lewis BC; Department of Clinical Pharmacology (N.C., L.K., B.C.L., P.I.M., J.O.M.) and Flinders Centre for Innovation in Cancer (B.C.L., P.I.M., J.O.M.), Flinders University College of Medicine and Public Health, Adelaide, Australia.
  • Mackenzie PI; Department of Clinical Pharmacology (N.C., L.K., B.C.L., P.I.M., J.O.M.) and Flinders Centre for Innovation in Cancer (B.C.L., P.I.M., J.O.M.), Flinders University College of Medicine and Public Health, Adelaide, Australia.
  • Miners JO; Department of Clinical Pharmacology (N.C., L.K., B.C.L., P.I.M., J.O.M.) and Flinders Centre for Innovation in Cancer (B.C.L., P.I.M., J.O.M.), Flinders University College of Medicine and Public Health, Adelaide, Australia john.miners@flinders.edu.au.
Drug Metab Dispos ; 47(3): 271-278, 2019 03.
Article in En | MEDLINE | ID: mdl-30541877
Accumulating evidence indicates that several human UDP-glucuronosyltransferase (UGT) enzymes catalyze both glucuronidation and glucosidation reactions. Baculovirus-infected insect cells [Trichoplusia ni and Spodoptera frugiperda (Sf9)] are used widely for the expression of recombinant human UGT enzymes. Following the observation that control Supersomes (c-SUP) express a native enzyme capable of glucosidating morphine, we characterized the glucosidation of a series of aglycones with a hydroxyl (aliphatic or phenolic), carboxylic acid, or amine functional group by c-SUP and membranes from uninfected Sf9 cells. Although both enzyme sources glucosidated the phenolic substrates investigated, albeit with differing activities, differences were observed in the selectivities of the native UDP-glucosyltransferases toward aliphatic alcohols, carboxylic acids, and amines. For example, zidovudine was solely glucosidated by c-SUP. By contrast, c-SUP lacked activity toward the amines lamotrigine and trifluoperazine and did not form the acyl glucoside of mycophenolic acid, reactions all catalyzed by uninfected Sf9 membranes. Glucosidation intrinsic clearances were high for several substrates, notably 1-hydroxypyrene (∼1400-1900 µl/min⋅mg). The results underscore the importance of including control cell membranes in the investigation of drug and chemical glucosidation by UGT enzymes expressed in T. ni (High-Five) and Sf9 cells. In a coincident study, we observed that UGT1A5 expressed in Sf9, human embryonic kidney 293T, and COS7 cells lacked glucuronidation activity toward prototypic phenolic substrates. However, Sf9 cells expressing UGT1A5 glucosidated 1-hydroxypyrene with UDP-glucuronic acid as the cofactor, presumably due to the presence of UDP-glucose as an impurity. Artifactual glucosidation may explain, at least in part, a previous report of phenolic glucuronidation by UGT1A5.
Subject(s)

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Subcellular Fractions / Recombinant Proteins / Xenobiotics / Glucuronosyltransferase / Spodoptera Limits: Animals / Humans Language: En Journal: Drug Metab Dispos Journal subject: FARMACOLOGIA Year: 2019 Document type: Article Affiliation country: Australia Country of publication: United States

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Subcellular Fractions / Recombinant Proteins / Xenobiotics / Glucuronosyltransferase / Spodoptera Limits: Animals / Humans Language: En Journal: Drug Metab Dispos Journal subject: FARMACOLOGIA Year: 2019 Document type: Article Affiliation country: Australia Country of publication: United States