期刊
PROTEOMICS
卷 14, 期 12, 页码 1457-1463出版社
WILEY
DOI: 10.1002/pmic.201300487
关键词
Digestion; Immobilization of trypsin; Magnetic polydopamine-coated magnetic graphene nanocomposites; MALDI-TOF MS; Nanoproteomics
资金
- National Basic Research Priorities Program [2012CB910601, 2013CB911201]
- National Natural Science Foundation of China [21075022, 20875017, 21105016]
- Research Fund for the Doctoral Program of Higher Education of China [20110071110007, 20100071120053]
- Shanghai Municipal Natural Science Foundation [11ZR1403200]
- Shanghai Leading Academic Discipline Project [B109]
In this work, polydopamine-coated magnetic graphene (MG@PDA) nanocomposites were synthesized by a facile method. Trypsin was then directly immobilized on the surface of the nanocomposites through simple PDA chemistry with no need for introducing any other coupling groups. The as-made MG@PDA nanocomposites inherit not only the large surface area of graphene which makes them capable of immobilizing high amount of trypsin (up to 0.175 mg/mg), but also the good hydrophilicity of PDA which greatly improves their biocompatibility. Moreover, the strong magnetic responsibility makes them easy to be separated from the digested peptide solution when applying a magnetic field. The feasibility of the trypsin-immobilized MG@PDA (MG@PDA-trypsin) nanocomposites for protein digestion was investigated and the results indicated their high digestion efficiency in a short digestion time (10 min). In addition, the reusability and stability of the MG@PDA-trypsin nanocomposites were also tested in our work. To further confirm the efficiency of MG@PDA-trypsin nanocomposites for proteome analysis, they were applied to digest proteins extracted from skimmed milk, followed by nano RPLC-ESI-MS/MS analysis, and a total of 321 proteins were identified, much more than those obtained by 16-h in-solution digestion (264 proteins), indicating the great potential of MG@PDA-trypsin nanocomposites as the supports for high-throughput proteome study.
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