title: Detection of disulfide linkage by chemical derivatization and mass spectrometry.
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.
authors.affiliation: Rieveschl Laboratories for Mass Spectrometry, Department of Chemistry, University of Cincinnati, 312 College Dr, Cincinnati, OH, 45221, USA, balasual@ucmail.uc.edu.
authors.forename: Balasubrahmanyam
authors.initials: B
authors.lastname: Addepalli
mesh_terms: D000478:Alkylation; D002853:Chromatography, Liquid / Q000379:methods; D003545:Cysteine / Q000737:chemistry; D004220:Disulfides / Q000737:chemistry*; D013058:Mass Spectrometry / Q000379:methods; D010084:Oxidation-Reduction; D017510:Protein Folding; D011506:Proteins / Q000737:chemistry*
chemical_list: D004220:Disulfides; D011506:Proteins; D003545:Cysteine