Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA

TY - JOUR

T1 - Precision templated bottom-up multiprotein nanoassembly through defined click chemistry linkage to DNA

AU - Marth, Gabriella

AU - Hartley, Andrew

AU - Reddington, Samuel

AU - Sargisson, Lauren

AU - Parcollet, Marlene

AU - Dunn, Katherine Elizabeth

AU - Jones, Dafydd

AU - Stulz, Eugen

N1 - © 2017 American Chemical Society

PY - 2017/4/17

Y1 - 2017/4/17

N2 - 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.

AB - 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.

KW - DNA nanotechnology

KW - copper-free click chemistry

KW - energy transfer

KW - expanded genetic code

KW - origami

KW - precision assembly

KW - protein engineering

UR - http://www.scopus.com/inward/record.url?scp=85019939393&partnerID=8YFLogxK

U2 - 10.1021/acsnano.7b01711

DO - 10.1021/acsnano.7b01711

M3 - Article

SN - 1936-0851

VL - 11

SP - 5003

EP - 5010

JO - ACS Nano

JF - ACS Nano

IS - 5

ER -