Using protein dimers to maximize the protein hybridization efficiency with multisite DNA origami scaffolds

Vikash Verma, Leena Mallik, Rizal Hariadi, Sivaraj Sivaramakrishnan, Georgios Skiniotis, Ajit P. Joglekar

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

DNA origami provides a versatile platform for conducting 'architecture-function' analysis to determine how the nanoscale organization of multiple copies of a protein component within a multi-protein machine affects its overall function. Such analysis requires that the copy number of protein molecules bound to the origami scaffold exactly matches the desired number, and that it is uniform over an entire scaffold population. This requirement is challenging to satisfy for origami scaffolds with many protein hybridization sites, because it requires the successful completion of multiple, independent hybridization reactions. Here, we show that a cleavable dimerization domain on the hybridizing protein can be used to multiplex hybridization reactions on an origami scaffold. This strategy yields nearly 100% hybridization efficiency on a 6-site scaffold even when using low protein concentration and short incubation time. It can also be developed further to enable reliable patterning of a large number of molecules on DNA origami for architecture-function analysis.

Original languageEnglish (US)
Article numbere0137125
JournalPLoS One
Volume10
Issue number9
DOIs
StatePublished - Sep 8 2015
Externally publishedYes

Fingerprint

Scaffolds
Dimers
hybridization
DNA
Proteins
proteins
Molecules
dimerization
Dimerization
Population

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Using protein dimers to maximize the protein hybridization efficiency with multisite DNA origami scaffolds. / Verma, Vikash; Mallik, Leena; Hariadi, Rizal; Sivaramakrishnan, Sivaraj; Skiniotis, Georgios; Joglekar, Ajit P.

In: PLoS One, Vol. 10, No. 9, e0137125, 08.09.2015.

Research output: Contribution to journalArticle

Verma, Vikash ; Mallik, Leena ; Hariadi, Rizal ; Sivaramakrishnan, Sivaraj ; Skiniotis, Georgios ; Joglekar, Ajit P. / Using protein dimers to maximize the protein hybridization efficiency with multisite DNA origami scaffolds. In: PLoS One. 2015 ; Vol. 10, No. 9.
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