Integration of online digestion and electrolytic reduction with mass spectrometry for rapid disulfide-containing protein structural analysis.

ABSTRACT: Bottom-up structural analysis of disulfide-bond containing proteins usually involves time-consuming offline enzymatic digestion, chemical reduction and thiol protection prior to mass spectrometric detection, which takes many hours. This paper presents an expedited bottom-up approach, employing desorption electrospray ionization-mass spectrometry (DESI-MS) coupled with online pepsin digestion and online electrochemical reduction of disulfide bonds. Peptides are generated in high digestion yield as its precursor protein in acidic aqueous solution flows through a pepsin column, which can undergo direct electrolysis. The electrolytic behaviors of peptides, as online monitored by DESI-MS, suggest the presence or absence of disulfide bonds in the peptides, and also provide information to relate

SUBMITTER: Zheng Q 

PROVIDER: S-EPMC4240030 | biostudies-literature | 2013 Nov

REPOSITORIES: biostudies-literature