Diagonal Electrophoresis for Detecting Disulfide Bridges
- 59 Downloads
Methods for identifying disulfide bridges have routinely employed “diagonal” procedures using two-dimensional paper or thin-layer electrophoresis. This essentially utilizes the difference in electrophoretic mobility of peptides containing either cysteine or cystine in a disulfide link, before and after oxidation with performic acid. It was first described by Brown and Hartley (1). Peptides unaltered by the performic acid oxidation have the same mobility in both dimensions and, therefore, lie on a diagonal. After oxidation, peptides that contain cysteine or were previously covalently linked produce one or two spots off the diagonal, respectively. This method has also been adapted for HPLC methodology and is discussed in Chapter 80.
KeywordsCyanogen Bromide Electrophoresis Buffer Paper Electrophoresis Xylene Cyanol Performic Acid
- 3.Creighton, T. E. (1983) Disulphide bonds between cysteine residues, in Protein Structure—a Practical Approach (Rickwood, D. and Hames B. D., eds.), IRL, Oxford, UK, pp. 155–167.Google Scholar