title: Synthesis and use of an isoform-specific affinity matrix in the purification of glutathione S-transferases from the housefly, Musca domestica (L.).
abstract: Glutathione may be linked to an agarose matrix which has been activated by treatment with epichlorhydrin. The resulting resin displayed group selectivity for the glutathione S-transferases of the housefly Musca domestica (L). The isoenzymes of low isoelectric point, which have little activity with substrates other than 1-chloro-2,4-dinitrobenzene, bound strongly to this matrix and were eluted with 10 mM glutathione at pH 7.4. On the other hand, the group of isoenzymes of higher isoelectric point, showing activity with other substrates such as 3,4-dichloronitrobenzene, did not bind. These isoenzymes did bind to a sulfobromophthalein-glutathione conjugate immobilized on agarose and could be eluted with 5 mM sulfobromophthalein at pH 7.4. The immobilized glutathione resin bound rat liver glutathione S-transferase subunits from all three molecular weight classes.
authors: School of Biological Science, Victoria University of Wellington, New Zealand.
authors: A G
authors: AG
authors: Clark
authors: S N
authors: SN
authors: Marshall
authors: A R
authors: AR
authors: Qureshi
mesh_terms: D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods*; D005978:Glutathione; D005982:Glutathione Transferase / Q000737:chemistry / Q000302:isolation & purification*; D006793:Houseflies / Q000201:enzymology*; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000302:isolation & purification; D008970:Molecular Weight; D010635:Phenolphthaleins; D012117:Resins, Synthetic / Q000138:chemical synthesis
chemical_list: D007527:Isoenzymes; D010635:Phenolphthaleins; D012117:Resins, Synthetic; D005982:Glutathione Transferase; D005978:Glutathione