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Alternating current‐assisted on‐plate proteolysis for MALDI‐TOF MS peptide mapping
Author(s) -
Wang Sheng,
Wei Bangguo,
Yang Pengyuan,
Chen Gang
Publication year - 2008
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200800401
Subject(s) - proteolysis , trypsin , peptide , chemistry , chromatography , matrix assisted laser desorption/ionization , cytochrome c , digestion (alchemy) , biochemistry , enzyme , desorption , adsorption , organic chemistry , mitochondrion
In this report, alternating current‐assisted on‐plate proteolysis has been developed for rapid peptide mapping. Protein solutions containing trypsin were allowed to digest directly on the spots of a stainless steel MALDI plate with the assistance of low‐voltage alternating current electricity. Alternating current (AC) was allowed to pass through the protein solutions via the MALDI plate and a platinum disc electrode. The feasibility and performance of the novel proteolysis approach were investigated by the digestion of BSA and cytochrome c (Cyt‐ c ). It was demonstrated that AC substantially enhanced the efficiency of proteolysis and the digestion time was significantly reduced to 5 min. The digests were identified by MALDI‐TOF MS with sequence coverages of 42% (BSA) and 77% (Cyt‐ c ) that were comparable to those obtained by using conventional in‐solution tryptic digestion. The present proteolysis strategy is simple and efficient, offering great promise for MALDI‐TOF MS peptide mapping.