Translocation of double-stranded DNA through membrane-adapted phi29 motor protein nanopores.
Biological pores have been used to study the transport of DNA and other molecules, but most pores have channels that allow only the movement of small molecules and single-stranded DNA and RNA. The bacteriophage phi29 DNA-packaging motor, which allows double-stranded DNA to enter the virus during maturation and exit during an infection, contains a connector protein with a channel that is between 3.6 and 6 nm wide. Here we show that a modified version of this connector protein, when reconstituted into liposomes and inserted into planar lipid bilayers, allows the translocation of double-stranded DNA. The measured conductance of a single connector channel was 4.8 nS in 1 M KCl. This engineered and membrane-adapted phage connector is expected to have applications in microelectromechanical sensing, microreactors, gene delivery, drug loading and DNA sequencing.
Wendell
David
D
Department of Biomedical Engineering, College of Engineering and College of Medicine, University of Cincinnati, Cincinnati, OH 45267, USA.
Jing
Peng
P
Geng
Jia
J
Subramaniam
Varuni
V
Lee
Tae Jin
TJ
Montemagno
Carlo
C
Guo
Peixuan
P
D001435:Bacteriophages / Q000378:metabolism*; D001692:Biological Transport; D036022:Capsid Proteins / Q000737:chemistry / Q000378:metabolism*; D004247:DNA / Q000378:metabolism*; D042003:DNA Packaging; D004279:DNA, Viral / Q000378:metabolism*; D015640:Ion Channel Gating; D008051:Lipid Bilayers / Q000378:metabolism*; D008081:Liposomes / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000737:chemistry / Q000378:metabolism*; D049329:Nanostructures / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D016062:Porosity
D036022:Capsid Proteins; D004279:DNA, Viral; D008051:Lipid Bilayers; D008081:Liposomes; D020409:Molecular Motor Proteins; C097975:portal protein, bacteriophage phi29; D004247:DNA