title: Chymotryptic proteolysis accelerated by alternating current for MALDI-TOF-MS peptide mapping.
abstract: Alternating current (AC) has been employed to enhance the efficiency of chymotryptic proteolysis for peptide mapping. It was allowed to flow through the mixture solution of proteins and chymotrypsin via a pair of platinum wire electrodes. Bovine serum albumin (BSA) and cytochrome c (Cyt-c) were digested by the novel proteolysis approach to demonstrate its feasibility and performance. The results indicated that AC significantly accelerated in-solution chymotryptic proteolysis and the digestion time was substantially reduced to 5 min. The digests were identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with sequence coverages of 46% (BSA) and 90% (Cyt-c) that were much better than those obtained by using 12-h conventional in-solution chymotryptic proteolysis. In addition, AC-assisted chymotryptic proteolysis was employed to digest human serum to demonstrate its suitability to complex protein sample. The present proteolysis strategy is simple and efficient and will find a wide range of applications in proteomic research.
authors.affiliation: Department of Chemistry, School of Pharmacy, Fudan University, Shanghai, China.
authors.forename: Sheng
authors.initials: S
authors.lastname: Wang
authors.forename: Ting
authors.initials: T
authors.lastname: Liu
authors.forename: Luyan
authors.initials: L
authors.lastname: Zhang
authors.forename: Gang
authors.initials: G
authors.lastname: Chen
authors.forename: Pengyuan
authors.initials: P
authors.lastname: Yang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D002918:Chymotrypsin / Q000737:chemistry*; D045304:Cytochromes c / Q000032:analysis; D004560:Electricity; D005240:Feasibility Studies; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D012710:Serum Albumin, Bovine / Q000032:analysis; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D012710:Serum Albumin, Bovine; D045304:Cytochromes c; D002918:Chymotrypsin