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Self-assembly and modular functionalization of three-dimensional crystals from oppositely charged proteins.

Liljeström V, Mikkilä J, Kostiainen MA - Nat Commun (2014)

Bottom Line: Well-developed, especially DNA-based, methods for their preparation exist, yet most techniques concentrate on molecular and synthetic nanoparticle systems in non-biocompatible environment.Here we describe the self-assembly and characterization of binary solids that consist of crystalline arrays of native biomacromolecules.Importantly, the whole preparation process takes place at room temperature in a mild aqueous medium allowing the processing of delicate biological building blocks into ordered structures with lattice constants in the nanometre range.

View Article: PubMed Central - PubMed

Affiliation: 1] Biohybrid Materials Group, Department of Biotechnology and Chemical Technology, Aalto University, 00076 Aalto, Finland [2] Molecular Materials Group, Department of Applied Physics, Aalto University, 00076 Aalto, Finland.

ABSTRACT
Multicomponent crystals and nanoparticle superlattices are a powerful approach to integrate different materials into ordered nanostructures. Well-developed, especially DNA-based, methods for their preparation exist, yet most techniques concentrate on molecular and synthetic nanoparticle systems in non-biocompatible environment. Here we describe the self-assembly and characterization of binary solids that consist of crystalline arrays of native biomacromolecules. We electrostatically assembled cowpea chlorotic mottle virus particles and avidin proteins into heterogeneous crystals, where the virus particles adopt a non-close-packed body-centred cubic arrangement held together by avidin. Importantly, the whole preparation process takes place at room temperature in a mild aqueous medium allowing the processing of delicate biological building blocks into ordered structures with lattice constants in the nanometre range. Furthermore, the use of avidin-biotin interaction allows highly selective pre- or post-functionalization of the protein crystals in a modular way with different types of functional units, such as fluorescent dyes, enzymes and plasmonic nanoparticles.

No MeSH data available.


Related in: MedlinePlus

Functionalization with enzymes.Biotinylated horseradish peroxidase (B-HRP, a) catalyses the one-electron oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to a blue-coloured charge transfer complex (b). Progress curves (c) and the derived initial velocities (V0, d) show that the crystals have a high catalytic activity (V0 >0.1 μM s−1) after functionalization.
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f7: Functionalization with enzymes.Biotinylated horseradish peroxidase (B-HRP, a) catalyses the one-electron oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to a blue-coloured charge transfer complex (b). Progress curves (c) and the derived initial velocities (V0, d) show that the crystals have a high catalytic activity (V0 >0.1 μM s−1) after functionalization.

Mentions: Crystalline arrays consisting of solely protein-based subunits offer an attractive platform for the creation of novel multifunctional biomaterials. We used B-HRP to functionalize the CCMV–avidin crystals with active enzymes (Fig. 7a). HRP catalyses the one-electron oxidation of 3,3′,5,5′-tetramethylbenzidine in the presence of hydrogen peroxide (Fig. 7b), which allows straightforward spectrophotometric quantitation (Supplementary Note 1). Structural characterization of crystals pre- or post functionalized with B-HRP shows a bcc crystal structure before and after the enzymatic reaction (Supplementary Fig. 4). Before washing, the functionalized crystals (irrespective of functionalization method) show high enzymatic activity, whereas the separated supernatant shows only low residual activity (Fig. 7c; Supplementary Fig. 5 and Supplementary Tables 2 and 3). For washed samples, the initial velocity (V0) for the reaction in the case of functionalized crystals (0.13−0.15 μM s−1) is ~2 orders of magnitude higher compared with the residual-free enzymes in the solution (~0.007 μM s−1; Fig. 7d), demonstrating that active enzymes can be efficiently loaded to and concentrated by the crystalline assemblies.


Self-assembly and modular functionalization of three-dimensional crystals from oppositely charged proteins.

Liljeström V, Mikkilä J, Kostiainen MA - Nat Commun (2014)

Functionalization with enzymes.Biotinylated horseradish peroxidase (B-HRP, a) catalyses the one-electron oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to a blue-coloured charge transfer complex (b). Progress curves (c) and the derived initial velocities (V0, d) show that the crystals have a high catalytic activity (V0 >0.1 μM s−1) after functionalization.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4109007&req=5

f7: Functionalization with enzymes.Biotinylated horseradish peroxidase (B-HRP, a) catalyses the one-electron oxidation of 3,3',5,5'-tetramethylbenzidine (TMB) to a blue-coloured charge transfer complex (b). Progress curves (c) and the derived initial velocities (V0, d) show that the crystals have a high catalytic activity (V0 >0.1 μM s−1) after functionalization.
Mentions: Crystalline arrays consisting of solely protein-based subunits offer an attractive platform for the creation of novel multifunctional biomaterials. We used B-HRP to functionalize the CCMV–avidin crystals with active enzymes (Fig. 7a). HRP catalyses the one-electron oxidation of 3,3′,5,5′-tetramethylbenzidine in the presence of hydrogen peroxide (Fig. 7b), which allows straightforward spectrophotometric quantitation (Supplementary Note 1). Structural characterization of crystals pre- or post functionalized with B-HRP shows a bcc crystal structure before and after the enzymatic reaction (Supplementary Fig. 4). Before washing, the functionalized crystals (irrespective of functionalization method) show high enzymatic activity, whereas the separated supernatant shows only low residual activity (Fig. 7c; Supplementary Fig. 5 and Supplementary Tables 2 and 3). For washed samples, the initial velocity (V0) for the reaction in the case of functionalized crystals (0.13−0.15 μM s−1) is ~2 orders of magnitude higher compared with the residual-free enzymes in the solution (~0.007 μM s−1; Fig. 7d), demonstrating that active enzymes can be efficiently loaded to and concentrated by the crystalline assemblies.

Bottom Line: Well-developed, especially DNA-based, methods for their preparation exist, yet most techniques concentrate on molecular and synthetic nanoparticle systems in non-biocompatible environment.Here we describe the self-assembly and characterization of binary solids that consist of crystalline arrays of native biomacromolecules.Importantly, the whole preparation process takes place at room temperature in a mild aqueous medium allowing the processing of delicate biological building blocks into ordered structures with lattice constants in the nanometre range.

View Article: PubMed Central - PubMed

Affiliation: 1] Biohybrid Materials Group, Department of Biotechnology and Chemical Technology, Aalto University, 00076 Aalto, Finland [2] Molecular Materials Group, Department of Applied Physics, Aalto University, 00076 Aalto, Finland.

ABSTRACT
Multicomponent crystals and nanoparticle superlattices are a powerful approach to integrate different materials into ordered nanostructures. Well-developed, especially DNA-based, methods for their preparation exist, yet most techniques concentrate on molecular and synthetic nanoparticle systems in non-biocompatible environment. Here we describe the self-assembly and characterization of binary solids that consist of crystalline arrays of native biomacromolecules. We electrostatically assembled cowpea chlorotic mottle virus particles and avidin proteins into heterogeneous crystals, where the virus particles adopt a non-close-packed body-centred cubic arrangement held together by avidin. Importantly, the whole preparation process takes place at room temperature in a mild aqueous medium allowing the processing of delicate biological building blocks into ordered structures with lattice constants in the nanometre range. Furthermore, the use of avidin-biotin interaction allows highly selective pre- or post-functionalization of the protein crystals in a modular way with different types of functional units, such as fluorescent dyes, enzymes and plasmonic nanoparticles.

No MeSH data available.


Related in: MedlinePlus