Chymotryptic proteolysis accelerated by alternating current for MALDI-TOF-MS peptide mapping

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• 作者：Sheng Wang ; Ting Liu ; Luyan Zhang ; Gang Chen ; Pengyuan Yang
• 关键词：Alternating current ; Proteolysis ; Mass spectrometry ; Protein ; Chymotrypsin
• 刊名：Journal of Proteomics
• 出版年：2009
• 出版时间：2 May 2009
• 年：2009
• 卷：72
• 期：4
• 页码：640-647
• 全文大小：873 K

文摘

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.