Abstract
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.
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References
Brown, J. R., and Hartley, B. S. (1966) Location of disulphide bridges by diagonal paper electrophoresis Biochem. J. 101, 214–228.
Michl, H. (1951) Paper electrophoresis at potential differences of 50 volts per centimetre. Monatschr. Chem. 82, 489–493.
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.
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© 1996 Humana Press Inc., Totowa, NJ
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Aitken, A., Learmonth, M. (1996). Diagonal Electrophoresis for Detecting Disulfide Bridges. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-60327-259-9_81
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DOI: https://doi.org/10.1007/978-1-60327-259-9_81
Publisher Name: Humana Press
Print ISBN: 978-0-89603-338-2
Online ISBN: 978-1-60327-259-9
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