Anomalous electrophoretic behaviour of the glutathione S-transferase Ya and Yk subunits isolated from man and rodents. A potential pitfall for nomenclature.
GSH S-transferases are dimeric enzymes. The subunits in the rat are resolved into six types, designated Yf, Yk, Ya, Yn, Yb and Yc, by discontinuous SDS/polyacrylamide-gel electrophoresis [Hayes (1986) Biochem. J. 233, 789-798]. The relative electrophoretic mobility of the Ya and Yk subunits is dependent on the amount of cross-linker (NN'-methylenebisacrylamide) in the resolving gel. At low degrees of cross-linking, CBis 0.6% (w/w), the Yk and Ya subunits possess a faster anodal mobility than do the Yf, Yn, Yb and Yc subunits (i.e. order of mobility Yk greater than Ya greater than Yf greater than Yn greater than Yb greater than Yc), whereas at higher degrees of cross-linking, CBis 5.0% (w/w), Yf subunits possess the fastest mobility (i.e. order of mobility Yf greater than Yk greater than or equal to Yn greater than Yb greater than or equal to Ya greater than Yc). Resolving gels that contain low concentrations of cross-linker [CBis 0.6% (w/w)] allow the resolution of a hitherto unrecognized polypeptide that is isolated by S-hexyl-GSH-Sepharose affinity chromatography. This new polypeptide, which we have designated Yb, is normally obscured by the main Yb band in resolving gels that comprise concentrations of cross-linker of at least CBis 1.6% (w/w). The Ya- and Yb-type subunits in guinea pig, mouse, hamster and man were identified by immuno-blotting and their apparent Mr values in different electrophoresis systems were determined. The Ya subunits in all species studied possess a variable cross-linker-dependent mobility during electrophoresis. Since the transferase subunits are currently classified according to their mobilities during SDS/polyacrylamide-gel electrophoresis, it is apparent that the variable electrophoretic behaviour of the Ya and Yk subunits may lead to the mis-identification of enzymes.
D000178:Acrylamides; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D005982:Glutathione Transferase / Q000302:isolation & purification*; D006801:Humans; D007122:Immunoelectrophoresis; D007527:Isoenzymes / Q000302:isolation & purification*; D008099:Liver / Q000032:analysis; D008297:Male; D010446:Peptide Fragments / Q000302:isolation & purification; D051381:Rats; D011919:Rats, Inbred Strains; D013045:Species Specificity; D009626:Terminology as Topic
D000178:Acrylamides; D007527:Isoenzymes; D010446:Peptide Fragments; D005982:Glutathione Transferase; C021221:N,N'-methylenebisacrylamide