Precision
Templated Bottom-Up Multiprotein Nanoassembly
through Defined Click Chemistry Linkage to DNA
Version 2 2017-05-22, 19:03
Version 1 2017-04-20, 14:27
Posted on 2017-05-22 - 19:03
We
demonstrate an approach that allows attachment of single-stranded
DNA (ssDNA) to a defined residue in a protein of interest (POI) so
as to provide optimal and well-defined multicomponent assemblies.
Using an expanded genetic code system, azido-phenylalanine (azF) was
incorporated at defined residue positions in each POI; copper-free
click chemistry was used to attach exactly one ssDNA at precisely
defined residues. By choosing an appropriate residue, ssDNA conjugation
had minimal impact on protein function, even when attached close to
active sites. The protein-ssDNA conjugates were used to (i) assemble
double-stranded DNA systems with optimal communication (energy transfer)
between normally separate groups and (ii) generate multicomponent
systems on DNA origami tiles, including those with enhanced enzyme
activity when bound to the tile. Our approach allows any potential
protein to be simply engineered to attach ssDNA or related biomolecules,
creating conjugates for designed and highly precise multiprotein nanoscale
assembly with tailored functionality.
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Marth, Gabriella; Hartley, Andrew M.; Reddington, Samuel C.; Sargisson, Lauren L.; Parcollet, Marlène; E. Dunn, Katherine; et al. (2017). Precision
Templated Bottom-Up Multiprotein Nanoassembly
through Defined Click Chemistry Linkage to DNA. ACS Publications. Collection. https://doi.org/10.1021/acsnano.7b01711
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AUTHORS (8)
GM
Gabriella Marth
AH
Andrew M. Hartley
SR
Samuel C. Reddington
LS
Lauren L. Sargisson
MP
Marlène Parcollet
KE
Katherine E. Dunn
DJ
D. Dafydd Jones
ES
Eugen Stulz
KEYWORDS
code systemssDNA conjugationmulticomponent systemsDefined Click Chemistry Linkagemulticomponent assembliessingle-stranded DNAmultiprotein nanoscale assemblyenergy transferPOIprotein functionapproachcopper-free click chemistryenzyme activityprecision Templated Bottom-Up Multiprotein NanoassemblyDNA origami tilesprotein-ssDNA conjugatesdouble-stranded DNA systemsresidue positions