Separation and localization on sodium dodecyl sulfate polyacrylamide gels of pig spleen lymphocyte plasma membrane proteins which bind 125I-labelled phytohemagglutinin.
The protein constituents of splenic pig lymphocytes derived from the plasma membranes, the endoplasmic reticulum or from a non-ionic (Nonidet P-40) detergent of whole cells have been separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, under reducing and non-reducing conditions. The binding of Phaseolus vulgaris (red kidney bean) phytohemagglutinin to these constituents has been studied by incubation of the labelled lectin with the undried electrophoretograms. Results show characteristic phytohemagglutinin-reactive components in each cell fraction, although the majority of the lectin-binding bands are common to the three fractions. Comparison of binding patterns for the reduced and non-reduced electrophoretograms does not reveal significant differences between the binding profiles. The components which bind the lectin in the largest amounts are located in the upper halves of the gels, which correspond to a molecular range of (75-250) . 10(3) daltons. Our data suggest that pig spleen lymphocyte plasma membranes contain at least 25-30 glycoproteins which can bind phytohemagglutinin. Iodine-labelled phytohemagglutinin binds to the vesicles prepared from lymphocyte plasma membranes and the Scatchard plot shows a non-linear upward concave curve.
D000818:Animals; D002462:Cell Membrane / Q000032:analysis; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D004721:Endoplasmic Reticulum; D008214:Lymphocytes / Q000032:analysis*; D008565:Membrane Proteins / Q000302:isolation & purification*; D008970:Molecular Weight; D017830:Octoxynol; D010835:Phytohemagglutinins; D011092:Polyethylene Glycols; D012967:Sodium Dodecyl Sulfate; D013154:Spleen / Q000032:analysis; D013552:Swine
D003902:Detergents; D008565:Membrane Proteins; D010835:Phytohemagglutinins; D012967:Sodium Dodecyl Sulfate; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40