Isoelectric-focusing properties and carbohydrate content of pea (Pisum sativum) legumin.
Legumin from pea (Pisum sativum) is a molecule made up of six pairs of subunits, each pair consisting of an ;acidic' subunit (mol.wt. about 40000) and a ;basic' subunit (mol.wt. about 20000) linked by one or more disulphide bonds. The heterogeneity of legumin has been investigated by isoelectric focusing; undissociated legumin could not be focused satisfactorily, but legumin subunits could be analysed under dissociating conditions. 8m-Urea was not found to be a satisfactory medium for isoelectric focusing of legumin, as the ;basic' subunits showed a shift in pI with time of incubation in urea. A new dissociating medium for isoelectric focusing, namely 50% (v/v) formamide, was used for analysis of legumin, which gave pI values of 5.0-5.3 for the ;acidic' subunits, and 8.3-8.7 for the ;basic' subunits. Both types of subunits were shown to be heterogeneous in charge and molecular weight by two-dimensional analysis employing isoelectric focusing in the first dimension and sodium dodecyl sulphate/polyacrylamide gel electrophoresis in the second. The ;basic' and ;acidic' subunits of legumin were separated on the preparative scale by ion-exchange chromatography in 50% formamide. Carbohydrate attached to the protein was investigated as a possible cause of the heterogeneity of legumin subunits. However, both a fluorescent-labelling technique and a sensitive radioactive-labelling technique failed to show any carbohydrate bound to legumin subunits, and it was concluded that legumin is not a glycoprotein.
D002241:Carbohydrates / Q000032:analysis*; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D007887:Fabaceae / Q000032:analysis; D005559:Formamides; D007525:Isoelectric Focusing; D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D010946:Plants, Medicinal; D014508:Urea; D000097793:Legumins
D002241:Carbohydrates; D005559:Formamides; D010940:Plant Proteins; D014674:Plant Proteins, Dietary; D014508:Urea; D000097793:Legumins