TY - JOUR
T1 - A nanoporous reactor for efficient proteolysis
AU - Qiao, Liang
AU - Liu, Yun
AU - Hudson, Sarah P.
AU - Yang, Pengyuan
AU - Magner, Edmond
AU - Liu, Baohong
PY - 2008
Y1 - 2008
N2 - 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.
AB - 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.
KW - Mass spectrometry
KW - Mesoporous materials
KW - Nanoreactors
KW - Nanostructures
KW - Proteolysis
UR - http://www.scopus.com/inward/record.url?scp=38049143633&partnerID=8YFLogxK
U2 - 10.1002/chem.200701102
DO - 10.1002/chem.200701102
M3 - Article
C2 - 17960551
AN - SCOPUS:38049143633
SN - 0947-6539
VL - 14
SP - 151
EP - 157
JO - Chemistry - A European Journal
JF - Chemistry - A European Journal
IS - 1
ER -