The effect of cross-links on the mobility of proteins in dodecyl sulphate-polyacrylamide gels.
The effect of reduction of intramolecular disulphide bridges on the mobility of proteins in 5% (w/v) polyacrylamide gels in the presence of sodium dodecyl sulphate was investigated. A series of polypeptide polymers, containing up to 68 intramolecular disulphide bridges, was prepared by cross-linking proteins of known structure with glutaraldehyde. These model polypeptides were denatured with heat, sodium dodecyl sulphate and urea, and their mobilities in sodium dodecyl sulphate-polyacrylamide gels compared before and after reduction with dithiothreitol. The mobilities of polypeptides containing no cystine were unaffected by reduction. However, reduction generally decreased the mobilities of polypeptides containing cystine; the extent of this decrease depended on the number of cystine residues originally present in the polypeptide polymer, and on the protein from which the latter was derived. In contrast with their higher oligomers, the monomer of lysozyme and the dimer of ribonuclease increased in mobility after reduction. The reduced polypeptide oligomers formed by reaction with glutaraldehyde were generally found to migrate at a rate significantly faster than was expected from their calculated molecular weights. It was concluded that the use of unreduced proteins and protein aggregates for molecular-weight measurements by the sodium dodecyl sulphate-polyacrylamide-gel method may give erroneous estimates of the molecular weight of any protein being investigated.
D000179:Acrylates; D000681:Amylases; D004586:Electrophoresis; D005233:Fatty Alcohols; D005782:Gels; D005934:Glucagon; D006454:Hemoglobins; D006358:Hot Temperature; D007782:Lactoglobulins; D008970:Molecular Weight; D009113:Muramidase; D011485:Protein Binding; D011506:Proteins; D012260:Ribonucleases; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D013501:Surface-Active Agents
D000179:Acrylates; D005233:Fatty Alcohols; D005782:Gels; D006454:Hemoglobins; D007782:Lactoglobulins; D011506:Proteins; D013501:Surface-Active Agents; D012710:Serum Albumin, Bovine; D012967:Sodium Dodecyl Sulfate; D005934:Glucagon; D012260:Ribonucleases; D000681:Amylases; D009113:Muramidase