title: Cytochrome c aided resolution of Lupinus albus isoperoxidases in a cathodal polyacrylamide gel electrophoresis system.
abstract: In a cathodal polyacrylamide gel electrophoresis system, three distinct groups of isoperoxidases from Lupinus albus were found to achieve retention factors (rf) dependent on the quantity of sample applied onto the gel. The possibility of extract-derived substances weakly associating with peroxidase samples was investigated. Association of the putative agents survived dialysis against electrophoresis buffer with and without 2 M CaCl2 and freeze-thaw treatments. The addition of polyvinylpolypyrrolidone and polyethylene glycol to the homogenization buffer also proved ineffective in eliminating the variation in isoperoxidase rf although differences in the zymogram profiles of these samples were evident. The addition of spermine and cytochrome c to samples was found to increase the rf of some peroxidase bands. Electrophoresis of samples with cytochrome c resulted in the resolution of peroxidase groups to distinct bands at rf independent of the quantity of peroxidase applied. Control experiments indicate that this treatment did not introduce any detectable artifacts.
authors.affiliation: Centro de Tecnologia Química e Biológica, Oeiras, Portugal.
authors.forename: P
authors.initials: P
authors.lastname: Jackson
authors.forename: C P
authors.initials: CP
authors.lastname: Ricardo
mesh_terms: D002473:Cell Wall / Q000201:enzymology; D003574:Cytochrome c Group / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D010544:Peroxidases / Q000302:isolation & purification* / Q000378:metabolism; D010944:Plants / Q000201:enzymology*
chemical_list: D003574:Cytochrome c Group; D007527:Isoenzymes; D010544:Peroxidases