Isolation and characterisation of the class alpha, mu and pi glutathione transferases in LLC-PK1 and pig kidney.
Glutathione S-transferase (GST) isoenzymes from pig kidney cortex and LLC-PK1 (an established cell line derived from the pig proximal tubule) were purified by affinity chromatography, anionic and cationic chromatofocusing. Purification revealed nine isoenzymes in the pig kidney cortex and five isoenzymes in the LLC-PK1 cell line. SDS-polyacrylamide gel electrophoresis showed that the pig kidney cortex isoenzymes were homo- or heterodimeric; LLC-PK1 isoenzymes, however, were homodimeric. Isoenzymes from pig and LLC-PK1 showed a higher affinity towards glutathione. The isoenzymes were further characterised and divided into the different GST classes by studying specific inhibitors, specific substrates and immunological properties. Pig GSTs belong to class alpha, mu and pi. The GSTs in LLC-PK1 cells, on the other hand, belong to class pi and mu. The isoenzyme pattern in LLC-PK1 cells indicates the dedifferentiation of this particular cell line compared with the pig kidney cortex.
Bohets
H H
HH
I.H.E., Afdeling Toxicologie, Brussels, Belgium.
Nouwen
E J
EJ
De Broe
M E
ME
Dierickx
P J
PJ
D000818:Animals; D002846:Chromatography, Affinity; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005260:Female; D005982:Glutathione Transferase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000502:physiology*; D007526:Isoelectric Point; D007527:Isoenzymes / Q000737:chemistry / Q000378:metabolism; D007672:Kidney Cortex / Q000737:chemistry / Q000201:enzymology*; D007687:Kidney Tubules, Proximal / Q000737:chemistry / Q000166:cytology / Q000201:enzymology*; D018374:LLC-PK1 Cells; D008970:Molecular Weight; D013379:Substrate Specificity; D013552:Swine
D004791:Enzyme Inhibitors; D007527:Isoenzymes; D005982:Glutathione Transferase