Mesoporous silica nanoreactors for highly efficient proteolysis.
Protein digestion inside the nanoreactor channels of mesoporous silica (SBA-15) is reported, and evaluated by using peptide-mass mapping. Both proteases and substrates were efficiently captured within these biocompatible nanoreactors. After 10 minutes, the mass spectrum of the protein digests released from the mesoporous-silica-based nanoreactors revealed the presence of eight peptides covering 58% of the protein sequence with an intense signal (signal/noise ratio > 70). In comparison, the conventional overnight in-solution digestion of proteins under otherwise identical conditions generated only three peptides (27% sequence coverage). We propose that this order-of-magnitude increase in the proteolytic reaction rate is mainly attributed to two factors: substrate enrichment within mesoporous silica channels and enzyme immobilization. The surface properties and macrostructure of the mesoporous silica were studied to reveal their significant influence on proteolytic reactions.
Department of Chemistry, Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, Fudan University, Shanghai 200433, PR China.
Jie
J
Fan
Wenqing
W
Shui
Pengyuan
P
Yang
Xiaoyan
X
Wang
Yunmin
Y
Xu
Honghai
H
Wang
Xian
X
Chen
Dongyuan
D
Zhao
D006868:Hydrolysis; D013058:Mass Spectrometry; D008855:Microscopy, Electron, Scanning; D046529:Microscopy, Electron, Transmission; D036103:Nanotechnology; D012822:Silicon Dioxide / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D012822:Silicon Dioxide