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Construction of a high-performance magnetic enzyme nanosystem for rapid tryptic digestion.

Cheng G, Zheng SY - Sci Rep (2014)

Bottom Line: The results show that the magnetic enzyme nanosystem can digest the proteins in 30 minutes, and the results are comparable to conventional 12 hours in-solution digestion.Furthermore, the magnetic enzyme nanosystem is also effective in the digestion of low-concentration proteins, even at as low as 5 ng μL(-1) substrate concentration.Therefore, this work will be highly beneficial for the rapid digestion and identification of proteins in future proteomics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical engineering, The Pennsylvania State University, University Park, PA 16802, (USA).

ABSTRACT
A magnetic enzyme nanosystem have been designed and constructed by a polydopamine (PDA)-modification strategy. The magnetic enzyme nanosystem has well defined core-shell structure and a relatively high saturation magnetization (Ms) value of 48.3 emu g(-1). The magnetic enzyme system can realize rapid, efficient and reusable tryptic digestion of proteins by taking advantage of its magnetic core and biofunctional shell. Various standard proteins (e.g. cytochrome C (Cyt-C), myoglobin (MYO) and bovine serum albumin (BSA)) have been used to evaluate the effectiveness of the magnetic enzyme nanosystem. The results show that the magnetic enzyme nanosystem can digest the proteins in 30 minutes, and the results are comparable to conventional 12 hours in-solution digestion. Furthermore, the magnetic enzyme nanosystem is also effective in the digestion of low-concentration proteins, even at as low as 5 ng μL(-1) substrate concentration. Importantly, the system can be reused several times, and has excellent stability for storage. Therefore, this work will be highly beneficial for the rapid digestion and identification of proteins in future proteomics.

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Schematic illustration of (a) the synthesis strategy for magnetic nanoreactor and (b) application as a multiplatform in rapid and recyclable digestion of proteins.
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f1: Schematic illustration of (a) the synthesis strategy for magnetic nanoreactor and (b) application as a multiplatform in rapid and recyclable digestion of proteins.

Mentions: Inspired by the bio-adhesion properties of marine mussels, polydopamine, a novel coating material, has sparked research interest and been moved into the spotlight3132. It can be easily coated on various inorganic and organic substrates with controllable film thickness and high stability33. More importantly, the chemical versatility, excellent dispersibility and extraordinary biocompatibility of polydopamine enable it to be a platform for diverse secondary reactions in biological applications (e.g. biomineralization, cell encapsulation, biomolecule immobilization)343536. Herein, we propose to construct a magnetic enzyme nanosystem (denoted as MEN) via a PDA-modification strategy for the rapid digestion of proteins. As shown in Figure 1a, the Fe3O4 magnetic particles were coated with a layer of polydopamine after being dispersed in an alkaline solution of dopamine hydrochloride, after which the trypsin molecules can be easily immobilized on the surface of the magnetic particles by simply mixing under mild conditions. The composite nanosystem combines the merits of the easy separation of Fe3O4 particles and the specific activity of trypsin functionalized polymer shells. They show the following advantages: (1) a polymer shell with a high density of functional groups would ensure the large binding capacity of trypsin molecules; (2) the large exposed surface area allows rapid mass transfer and diffusion of reactants and products; (3) the stability of the magnetic enzyme system under typical proteolytic digestion conditions and fast magnetic separation enables the regeneration and reuse of the nanosystem.


Construction of a high-performance magnetic enzyme nanosystem for rapid tryptic digestion.

Cheng G, Zheng SY - Sci Rep (2014)

Schematic illustration of (a) the synthesis strategy for magnetic nanoreactor and (b) application as a multiplatform in rapid and recyclable digestion of proteins.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4221791&req=5

f1: Schematic illustration of (a) the synthesis strategy for magnetic nanoreactor and (b) application as a multiplatform in rapid and recyclable digestion of proteins.
Mentions: Inspired by the bio-adhesion properties of marine mussels, polydopamine, a novel coating material, has sparked research interest and been moved into the spotlight3132. It can be easily coated on various inorganic and organic substrates with controllable film thickness and high stability33. More importantly, the chemical versatility, excellent dispersibility and extraordinary biocompatibility of polydopamine enable it to be a platform for diverse secondary reactions in biological applications (e.g. biomineralization, cell encapsulation, biomolecule immobilization)343536. Herein, we propose to construct a magnetic enzyme nanosystem (denoted as MEN) via a PDA-modification strategy for the rapid digestion of proteins. As shown in Figure 1a, the Fe3O4 magnetic particles were coated with a layer of polydopamine after being dispersed in an alkaline solution of dopamine hydrochloride, after which the trypsin molecules can be easily immobilized on the surface of the magnetic particles by simply mixing under mild conditions. The composite nanosystem combines the merits of the easy separation of Fe3O4 particles and the specific activity of trypsin functionalized polymer shells. They show the following advantages: (1) a polymer shell with a high density of functional groups would ensure the large binding capacity of trypsin molecules; (2) the large exposed surface area allows rapid mass transfer and diffusion of reactants and products; (3) the stability of the magnetic enzyme system under typical proteolytic digestion conditions and fast magnetic separation enables the regeneration and reuse of the nanosystem.

Bottom Line: The results show that the magnetic enzyme nanosystem can digest the proteins in 30 minutes, and the results are comparable to conventional 12 hours in-solution digestion.Furthermore, the magnetic enzyme nanosystem is also effective in the digestion of low-concentration proteins, even at as low as 5 ng μL(-1) substrate concentration.Therefore, this work will be highly beneficial for the rapid digestion and identification of proteins in future proteomics.

View Article: PubMed Central - PubMed

Affiliation: Department of Biomedical engineering, The Pennsylvania State University, University Park, PA 16802, (USA).

ABSTRACT
A magnetic enzyme nanosystem have been designed and constructed by a polydopamine (PDA)-modification strategy. The magnetic enzyme nanosystem has well defined core-shell structure and a relatively high saturation magnetization (Ms) value of 48.3 emu g(-1). The magnetic enzyme system can realize rapid, efficient and reusable tryptic digestion of proteins by taking advantage of its magnetic core and biofunctional shell. Various standard proteins (e.g. cytochrome C (Cyt-C), myoglobin (MYO) and bovine serum albumin (BSA)) have been used to evaluate the effectiveness of the magnetic enzyme nanosystem. The results show that the magnetic enzyme nanosystem can digest the proteins in 30 minutes, and the results are comparable to conventional 12 hours in-solution digestion. Furthermore, the magnetic enzyme nanosystem is also effective in the digestion of low-concentration proteins, even at as low as 5 ng μL(-1) substrate concentration. Importantly, the system can be reused several times, and has excellent stability for storage. Therefore, this work will be highly beneficial for the rapid digestion and identification of proteins in future proteomics.

Show MeSH