Peptide mapping of heterogeneous protein samples.
A simple two-dimensional electrophoretic method for peptide mapping of heterogeneous protein samples is presented. The reduced and denatured proteins of the mixture are separated in a first dimension by sodium dodecyl sulfate-polyacrylamide slab gel electrophoresis. After completion of the electrophoresis, the whole gel lane is equilibrated in stacking gel buffer and is transferred at right angles onto a second slab gel. A protease solution is overlayed on the gel lane and a partial proteolysis of the proteins to be analyzed is performed during the stacking phase of the second electrophoresis. The second electrophoresis resolves the characteristic pattern of peptides of each individual protein as a series of spots located below the original position of the undigested protein. The peptide maps of the following samples are presented as examples: protein P23 and P23* of bacteriophage T4, membranes of Dictyostelium discoideum, membranes of human erythrocytes, and 35S-labeled proteins of D. discoideum synthesized in vivo or in a cell-free wheat germ extract. In complex samples, up to 20 individual proteins can be analyzed at once and a protein comprising only 1% of the total sample generates a clearly identifiable peptide pattern. Good reproducibility of the patterns obtained allows the comparison of samples of different origins.
D002462:Cell Membrane / Q000032:analysis; D003090:Coliphages / Q000032:analysis; D004023:Dictyostelium / Q000032:analysis; D004591:Electrophoresis, Polyacrylamide Gel; D004910:Erythrocyte Membrane / Q000032:analysis; D006801:Humans; D008565:Membrane Proteins; D010446:Peptide Fragments / Q000032:analysis*; D010447:Peptide Hydrolases; D010940:Plant Proteins; D011506:Proteins; D013211:Staphylococcus aureus; D014764:Viral Proteins
D008565:Membrane Proteins; D010446:Peptide Fragments; D010940:Plant Proteins; D011506:Proteins; D014764:Viral Proteins; D010447:Peptide Hydrolases