Macroporous materials as novel catalysts for efficient and controllable proteolysis.
A novel nanopore based digestion strategy has been developed by directly adding a macroporous material as catalyst to the conventional in-solution reaction system. Without increasing the enzyme or protein concentrations, this simple digestion approach exhibits high proteolysis efficiency and selectivity due to the in situ fast adsorption of both enzymes and proteins from bulk solution into the macropores of the catalysts, where the target substrates and enzymes are greatly concentrated and confined in the nanospace to realize a quick digestion. Based on the electrostatic interaction matching between the biomolecules and catalysts, selective extraction and digestion of proteins with different isoelectric points can be achieved by adjusting the surface charge of the catalysts. This nanoporous reaction system has been successfully applied to the analysis of a complex biological sample, where 293 proteins are identified, while only 100 proteins are obtained by the standard overnight in-solution digestion. The present nanospace confined digestion strategy will lead to promising advances not only in proteomics but also in other applications where enzymatic reactions are involved.
Qian
Kun
K
Department of Chemistry and Shanghai Key Laboratory of Molecular Catalysis and Innovative Materials, Institute of Biomedical Sciences, Fudan University, Shanghai 200433, P. R. China.
Wan
Jingjing
J
Qiao
Liang
L
Huang
Xiaodan
X
Tang
Jiawei
J
Wang
Yunhua
Y
Kong
Jilie
J
Yang
Pengyuan
P
Yu
Chengzhong
C
Liu
Baohong
B
D000818:Animals; D055162:Biocatalysis; D002417:Cattle; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism; D004800:Enzymes, Immobilized / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008277:Magnesium Oxide / Q000737:chemistry; D016062:Porosity; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity
D004798:Enzymes; D004800:Enzymes, Immobilized; D011506:Proteins; D008277:Magnesium Oxide