Detection of disulfide linkage by chemical derivatization and mass spectrometry
1Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, 312 College Dr, Cincinnati, OH, 45221, USA, balasual@ucmail.uc.edu.
Abstract:
The location of disulfide linkage(s) or status of unpaired cysteines is a critical structural feature required for the characterization of three-dimensional structure of a protein and for the correlation of protein structure-function relationships. Cysteine, with its reactive thiol group, can undergo enzymatic or oxidative posttranslational modification in response to changing redox conditions to signal a cascade of downstream reactions. In such a situation, it becomes even more critical to obtain the information on the pair of cysteines involved in such a redox switch operation. Here, a method involving chemical derivatization and liquid chromatography-mass spectrometry (LC-MS) is described to determine the cysteine residues involved in disulfide bond formation for a protein containing multiple cysteines in its sequence.
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