Abstract
Single molecules of DNA or RNA can be detected as they are driven through an alpha-hemolysin channel by an applied electric field. During translocation, nucleotides within the polynucleotide must pass through the channel pore in sequential, single-file order because the limiting diameter of the pore can accommodate only one strand of DNA or RNA at a time. Here we demonstrate that this nanopore behaves as a detector that can rapidly discriminate between pyrimidine and purine segments along an RNA molecule. Nanopore detection and characterization of single molecules represent a new method for directly reading information encoded in linear polymers, and are critical first steps toward direct sequencing of individual DNA and RNA molecules.
Keywords
Non-programmatic
MeSH Terms
Bacterial Toxins
Base Sequence
Biophysical Phenomena
Biophysics
DNA, Single-Stranded/chemistry
Hemolysin Proteins
Ion Channels
Lipid Bilayers
Models, Molecular
Nucleic Acid Conformation
Poly A/chemistry
Poly C/chemistry
Poly U/chemistry
RNA/chemistry
Chemicals
Bacterial Toxins
DNA, Single-Stranded
Hemolysin Proteins
Ion Channels
Lipid Bilayers
staphylococcal alpha-toxin
Poly A
Poly U
Poly C
RNA
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Akeson M
Department of Chemistry & Biochemistry, University of California, Santa Cruz, CA 95064, USA. makeson@chemistry ucsc.edu
Branton D
Kasianowicz J J
Brandin E
Deamer D W
References (15)
15 references, click to expand
-
Horizontal 'solvent-free' lipid bimolecular membranes with two-sided access can be formed and facilitate ion channel reconstitution.
Biochim Biophys Acta. 1995 Jun 14;1236(2):339-44
PMID: 7540872
-
Characterization of individual polynucleotide molecules using a membrane channel.
Proc Natl Acad Sci U S A. 1996 Nov 26;93(24):13770-3
PMID: 8943010
-
Circular dichroism of cytosine dinucleoside monophosphates containing arabinose, ribose, and deoxyribose.
Biochemistry. 1968 Nov;7(11):3836-41
PMID: 5722251
-
Single-stranded oligomers and polymers of cytidylic and 2'-deoxycytidylic acids: comparative optical rotatory studies.
Proc Natl Acad Sci U S A. 1967 Feb;57(2):423-30
PMID: 16591487
-
A structural model for the polyadenylic acid single helix.
J Mol Biol. 1975 Apr 25;93(4):529-34
PMID: 1142433
-
Adenylate oligomers in single- and double-strand conformation.
J Mol Biol. 1966 Feb;15(2):467-88
PMID: 5915178
-
Conformation of polyribouridylic acid in solution.
J Mol Biol. 1970 Jun 14;50(2):373-89
PMID: 5476918
-
THE HELICAL CONFORMATIONS OF POLYCYTIDYLIC ACID: STUDIES ON THE FORCES INVOLVED.
Biochemistry. 1964 Aug;3:1015-21
PMID: 14220659
-
A study of polyadenylic acid at neutral pH.
J Mol Biol. 1966 Feb;15(2):455-66
PMID: 5915177
-
Nucleotide rigidity.
Nature. 1976 Oct 14;263(5578):567-72
PMID: 980104
-
Structure of staphylococcal alpha-hemolysin, a heptameric transmembrane pore.
Science. 1996 Dec 13;274(5294):1859-66
PMID: 8943190
-
Structure of the single-stranded polyribonucleotide polycytidylic acid.
J Mol Biol. 1976 Sep 25;106(3):735-48
PMID: 10446
-
Optimizing planar lipid bilayer single-channel recordings for high resolution with rapid voltage steps.
Biophys J. 1990 Aug;58(2):289-97
PMID: 1698470
-
Patch clamp detection of transcription factor translocation along the nuclear pore complex channel.
J Membr Biol. 1995 Aug;146(3):253-61
PMID: 8568840
-
Counting polymers moving through a single ion channel.
Nature. 1994 Jul 28;370(6487):279-81
PMID: 7518571