Single‐Molecule Covalent Chemistry with Spatially Separated Reactants

Luchian, Tudor; Shin, Seong‐Ho; Bayley, Hagan

doi:10.1002/anie.200351313

Cited by 107 publications

(107 citation statements)

References 28 publications

(34 reference statements)

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“…For example, in stochastic sensing a wide variety of analytes can be detected at the single molecule level through the modulation of the current flowing through a single pore (29 -31). Further, the ␣HL pore has recently been used as a nanoreactor to examine chemical reactions at the single molecular level (32)(33)(34)(35). Accordingly, the ␣HL pore is emerging as a useful tool in both basic science and biotechnology (28,31).…”

Section: Lytic (Stage 3mentioning

confidence: 99%

Role of the Amino Latch of Staphylococcal α-Hemolysin in Pore Formation

Jayasinghe

Miles

Bayley

2006

Journal of Biological Chemistry

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Staphylococcal ␣-hemolysin (␣HL) is a ␤ barrel pore-forming toxin that is secreted by the bacterium as a water-soluble monomeric protein. Upon binding to susceptible cells, ␣HL assembles via an inactive prepore to form a water-filled homoheptameric transmembrane pore. The N terminus of ␣HL, which in the crystal structure of the fully assembled pore forms a latch between adjacent subunits, has been thought to play a vital role in the prepore to pore conversion. For example, the deletion of two N-terminal residues produced a completely inactive protein that was arrested in assembly at the prepore stage. In the present study, we have re-examined assembly with a comprehensive set of truncation mutants. Surprisingly, we found that after truncation of up to 17 amino acids, the ability of ␣HL to form functional pores was diminished, but still substantial. We then discovered that the mutation Ser 217 3 Asn, which was present in our original set of truncations but not in the new ones, promotes complete inactivation upon truncation of the N terminus. Therefore, the N terminus of ␣HL cannot be critical for the prepore to pore transformation as previously thought. Residue 217 is involved in the assembly process and must interact indirectly with the distant N terminus during the last step in pore formation. In addition, we provide evidence that an intact N terminus prevents the premature oligomerization of ␣HL monomers in solution.

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Section: Lytic (Stage 3mentioning

confidence: 99%

Role of the Amino Latch of Staphylococcal α-Hemolysin in Pore Formation

Jayasinghe

Miles

Bayley

2006

Journal of Biological Chemistry

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“…The identity and the concentration of the analyte could be revealed from its characteristic current signatures such as the event residence time, amplitude, frequency, and even the shape of the blockage. 19 In addition to its biosensing application, 20−29 nanopore sensing technology has been successfully applied to study a variety of other research areas, for example, covalent and noncovalent bonding interactions, 30,31 biomolecular folding and unfolding, 32−35 and enzyme kinetics. 36−38 It should be noted that nanopore biosensing is generally achieved by modifying the nanopore interior to introduce binding sites for molecular recognition of target analytes.…”

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confidence: 99%

Peptide-Mediated Nanopore Detection of Uranyl Ions in Aqueous Media

et al. 2017

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Uranium is one of the most common radioactive contaminants in the environment. As a major nuclear material in production, environmental samples (like soil and groundwater) can provide signatures on uranium production activity inside the facility. Thus, developing a new and portable analytical technology for uranium in aqueous media is significant not only for environmental monitoring, but also for nonproliferation. In this work, a label-free method for the detection of uranyl (UO22+) ions is developed by monitoring the translocation of a peptide probe in a nanopore. Based on the difference in the number of peptide events in the absence and presence of uranyl ions, nanomolar concentration of UO22+ ions could be detected in minutes. The method is highly selective; micromolar concentrations of Cd2+, Cu2+, Zn2+, Ni2+, Pb2+, Hg2+, Th4+, Mg2+, and Ca2+ would not interfere with the detection of UO22+ ions. In addition, simulated water samples were successfully analyzed.

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“…Along the same lines, it was possible to detect and characterize the short-lived intermediate 211 of the reduction of disulfide 212 with dithiothreitol 213 within a bioengineered a-hemolysin pore 214. 209 These pioneering examples from biological and bioengineered pores on regulation, medicinal applications and sensing give an excellent flavor of the fact that the perspectives in the field of synthetic ion channels and pores are extremely attractive and without limits. We look forward to reviewing the next four-year period, January 2004 to December 2007, with the highest expectations.…”

Section: Perspectivesmentioning

confidence: 99%