Preparation of Symmetrical and Unsymmetrical DNA–Protein Conjugates with DNA as a Molecular Scaffold

Tomkins, Justin M.; Nabbs, Brent K.; Barnes, Karen J.; Legido, Marta; Blacker, A. John; McKendry, Rachel A.; Abell, Chris

doi:10.1002/1439-7633(20010504)2:5<375::aid-cbic375>3.3.co;2-9

Cited by 4 publications

(6 citation statements)

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“…The potential of DNA as a building block in nanotechnology is recognized in its use in the creation of novel structures 1 and as a molecular scaffold. 2 The interaction between surface-immobilized DNA and free oligonucleotides in solution plays an important role in a wide range of array-based genetic diagnostic devices. 3 The density, organization, and properties of surface immobilized DNA have been studied by a range of techniques that include surface force, 4 quartz crystal microbalance (QCM), 5 surface plasmon resonance (SPR), 6 X-ray photoelectron spectroscopy (XPS), 7 neutron reflection, 8 and electrochemical response.…”

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confidence: 99%

Use of Atomic Force Microscopy for Making Addresses in DNA Coatings

et al. 2002

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Nanoscale DNA surfaces are manipulated by coating ultraflat gold with short thiolated single-stranded (ss) and double-stranded (ds) DNA. Hybridization and 6-mercapto-1-hexanol (MCH) are used to modulate the thickness of the layers, all the changes being monitored using atomic force microscopy (AFM) in situ. Short thiolated ssDNA forms irregular 3.5 nm thick coatings, which are converted into more uniform 6.0 nm thick layers upon hybridization with complementary DNA. Holes formed in a monolayer of short thiolated dsDNA can be partially filled with a longer length thiolated ssDNA, which then protrudes from the surface after hybridization with a complementary strand. MCH is used to modulate the depth of a ssDNA layer and improve hybridization to it.

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confidence: 99%

Use of Atomic Force Microscopy for Making Addresses in DNA Coatings

et al. 2002

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“…There are currently several methodologies used to link proteins to DNA. Proteins have been assembled onto DNA scaffolds through intervening adapter molecules such as streptavidin12, 15–20 or aptamers 21. 22 Alternately, direct covalent conjugation can be achieved by modification of cysteine or lysine residues23–26 or intein modification 11.…”

Section: Methodsmentioning

confidence: 99%

A Universal Method for the Preparation of Covalent Protein–DNA Conjugates for Use in Creating Protein Nanostructures

et al. 2007

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DNA has numerous attractive features as a scaffold for nanostructure assembly. Its rigidity, predictable structure, and assembly through complementary hybridization allow DNA to form nanoscale architectures such as cubes, [1] tetrahedra, [2] octahedra, [3,4] and 2D arrays. [5][6][7][8] By introducing proteins into DNA nanostructures, the recognition elements and functionalities that are inherent in proteins can be organized into nanostructured motifs. DNA-scaffolded protein assemblies have been used in immuno-PCR detection methods (PCR = polymerase chain reaction) [9][10][11] to arrange biocatalysts in a series for sequential reactions [12,13] and to organize other nanomaterials. [14] There are currently several methodologies used to link proteins to DNA. Proteins have been assembled onto DNA scaffolds through intervening adapter molecules such as streptavidin [12,[15][16][17][18][19][20] or aptamers. [21,22] Alternately, direct covalent conjugation can be achieved by modification of cysteine or lysine residues [23][24][25][26] or intein modification. [11,27,28] Niemyer and co-workers have employed these protein-DNA conjugates to form fluorescence resonant energy transfer (FRET) systems for use in nanobiotechnology. [29,30] Herein, we demonstrate a fusion-based strategy to regioselectively and covalently label proteins at the C terminus with single-stranded DNA. These protein-oligonucleotide chimeras were then spontaneously assembled into nanoarchitectures by complementary hybridization of the DNA. The covalent attachment strategy described herein yields a short and compact linkage between the protein and DNA molecule that allows for precise control over protein spacing and orientation in the final nanostructure.To achieve selective protein labeling, we use the enzyme protein farnesyltransferase (PFTase) to label a substrate protein containing a C-terminal tetrapeptide tag with an azide-modified isoprenoid diphosphate (1, Scheme 1).

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“…By thermal denaturation and rapid cooling, interesting finger ring-like DNA streptavin nanocircles were observed whose dimensions varied from 12 nm to 55 nm [44]. In another example, Tomkins et al fabricated nanoscale symmetrical and nonsymmetrical dumbbells using DNA as a molecular scaffold for streptavidin attachment [45]. Another interesting noncovalent binding of proteins with DNA is based on DNA aptamer-directed self-assembly [46].…”

Section: Dna and Proteinmentioning

confidence: 99%