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Proc Natl Acad Sci U S A. 2007 Jul 31;104(31):12646-50. Epub 2007 Jun 11.

Fueling protein DNA interactions inside porous nanocontainers.

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1
Department of Physics, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.

Erratum in

  • Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14878.

Abstract

Vesicle encapsulation offers a biologically relevant environment for many soluble proteins and nucleic acids and an optimal immobilization medium for single-molecule fluorescence assays. Furthermore, the confinement of biomolecules within small volumes opens up new avenues to unique experimental configurations. Nevertheless, the vesicles' impermeability, even toward ions and other small molecules such as ATP, hinders more general applications. We therefore developed methods to induce pores into vesicles. Porous vesicles were then used to modulate the interaction between Escherichia coli RecA proteins and ssDNA by changing the extravesicular nucleotides. Repetitive binding and dissociation of the same RecA filament on the DNA was observed with a rebinding rate two orders of magnitude greater than in the absence of confinement, suggesting a previously unreported nucleation pathway for RecA filament. This method provides a biofriendly and simple alternative to surface tethering that is ideal for the study of transient and weakly interacting biological complexes.

PMID:
17563361
PMCID:
PMC1937520
DOI:
10.1073/pnas.0610673104
[Indexed for MEDLINE]
Free PMC Article
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