Capillary electrophoresis of seed 2S albumins from Lupinus species.
Two modes of capillary electrophoresis (CE)--free-solution capillary zone electrophoresis (CZE) and sodium dodecyl sulfate capillary electrophoresis (SDS-CE) using a non-gel sieving matrix--have been developed for comparative analysis of low-molecular-mass 2S albumin isoforms from lupins. The albumin fraction and 2S albumins were separated in uncoated fused-silica capillary by CZE with 0.02 M phosphate buffer, pH 7.3, containing the sodium salt of phytic acid. The use of phytic acid (0.025 M) as buffer modifier and ion-pairing agent improved migration reproducibility, peak shape and separation efficiency. The reduced 2S albumins were separated by SDS-CE using a high concentration (0.3-0.5 M) mixture of tris(hydroxymethyl)aminomethane and borate buffers in uncoated fused-silica capillary. Of the various polymers used as non-gel sieving matrix, SDS-CE with a 10% dextran solution was found to be suitable for separation of 2S albumin polypeptides with molecular masses of 4,000-7,000 and 8,000-11,000. The addition of glycerol or ethylene glycol to the SDS separating buffer improved the resolution of polypeptides. The examined Lupinus species showed species-specific CZE and SDS-CE migration profiles of the 2S albumins.
Salmanowicz
B P
BP
Institute of Plant Genetics, Polish Academy of Sciences, Poznań. bsal@igrnov.igr.poznan.pl
D000418:Albumins / Q000302:isolation & purification*; D002021:Buffers; D019075:Electrophoresis, Capillary / Q000379:methods*; D007887:Fabaceae / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D010946:Plants, Medicinal; D015203:Reproducibility of Results; D012639:Seeds / Q000737:chemistry*; D013045:Species Specificity
D000418:Albumins; D002021:Buffers