A nanoporous reactor for efficient proteolysis

Liang Qiao, Yun Liu, Sarah P. Hudson, Pengyuan Yang, Edmond Magner, Baohong Liu

Research output: Contribution to journalArticlepeer-review

Abstract

A nanoreactor based on mesoporous silicates is described for efficient tryptic digestion of proteins within the mesochannels. Cyano-functionalized mcsoporous silicate (CNS), with an average pore diameter of 18 nm, is a good support for trypsin, with rapid in situ digestion of the model proteins, cytochrome c and myoglobin. The generated peptides were analyzed by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). Proteolysis by trypsin-CNS is much more efficient than in-solution digestion, which can be attributed to nanoscopic confinement and concentration enrichment of the substrate within the mesopores. Proteins at concentrations of 2 ng μL 1 were successfully identified after digestion for 20 min. A biological complex sample extracted from the cytoplasm of human liver tissue was digested by using the CNS-based reactor. Coupled with reverse-phase HPLC and MALDI-TOF MS/MS, 165 proteins were identified after standard protein data searching. This nanoreactor combines the advantages of short digestion time with retention of enzymatic activity, providing a promising way to advance the development of proteomics.

Original languageEnglish
Pages (from-to)151-157
Number of pages7
JournalChemistry - A European Journal
Volume14
Issue number1
DOIs
Publication statusPublished - 2008

Keywords

  • Mass spectrometry
  • Mesoporous materials
  • Nanoreactors
  • Nanostructures
  • Proteolysis

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