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Journal ArticleDOI

In singulo Biochemistry: When Less Is More

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TLDR
Some of the advantages of single-molecule methods over their bulk counterparts are discussed and it is argued that these advantages should help establish them as essential tools in the technical arsenal of the modern biochemist.
Abstract
It has been over one-and-a-half decades since methods of single-molecule detection and manipulation were first introduced in biochemical research. Since then, the application of these methods to an expanding variety of problems has grown at a vertiginous pace. While initially many of these experiments led more to confirmatory results than to new discoveries, today single-molecule methods are often the methods of choice to establish new mechanism-based results in biochemical research. Throughout this process, improvements in the sensitivity, versatility, and both spatial and temporal resolution of these techniques has occurred hand in hand with their applications. We discuss here some of the advantages of single-molecule methods over their bulk counterparts and argue that these advantages should help establish them as essential tools in the technical arsenal of the modern biochemist.

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Citations
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Sending signals dynamically.

TL;DR: The next stages in the signaling hierarchy—how multiple signals are integrated and how cellular signaling pathways are organized in space and time—present exciting challenges for the future, requiring bold multidisciplinary approaches.
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Covalent mechanochemistry: theoretical concepts and computational tools with applications to molecular nanomechanics.

TL;DR: From Macroscopic Milling to Bond-Selective Manipulation from macroscopic milling to bond-selected Manipulation is described.
Book ChapterDOI

Accurate Single-Molecule FRET Studies Using Multiparameter Fluorescence Detection

TL;DR: In this paper, the authors demonstrate that the simultaneous acquisition of a maximum of fluorescence parameters by multiparameter fluorescence detection (MFD) allows for a robust assessment of all possible artifacts arising from smFRET and offers unsurpassed capabilities regarding the identification and analysis of individual species present in a population of molecules.
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Single-molecule analysis using DNA origami.

TL;DR: Recent progress in the area of single-molecule analysis using DNA origami is summarized and the future directions of this research are discussed.
Journal ArticleDOI

Enzymatic oxygen scavenging for photostability without pH drop in single-molecule experiments.

TL;DR: Pyranose oxidase and catalase is presented as a novel enzymatic system to perform single-molecule experiments in pH-stable conditions at arbitrary buffer strength and enhanced stability allows the observation of bionanotechnological assemblies in aqueous environments under well-defined conditions for an extended time.
References
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Journal ArticleDOI

Nonequilibrium Equality for Free Energy Differences

TL;DR: In this paper, an expression for the equilibrium free energy difference between two configurations of a system, in terms of an ensemble of finite-time measurements of the work performed in parametrically switching from one configuration to the other, is derived.
Journal ArticleDOI

Reversible Unfolding of Individual Titin Immunoglobulin Domains by AFM

TL;DR: Single-molecule atomic force microscopy was used to investigate the mechanical properties of titin, the giant sarcomeric protein of striated muscle, and refolding of immunoglobulin domains was observed.
Journal ArticleDOI

Entropy production fluctuation theorem and the nonequilibrium work relation for free energy differences

TL;DR: A generalized version of the fluctuation theorem is derived for stochastic, microscopically reversible dynamics and this generalized theorem provides a succinct proof of the nonequilibrium work relation.
Journal ArticleDOI

Direct observation of kinesin stepping by optical trapping interferometry

TL;DR: It is found that kinesin moves with 8-nm steps, similar to biological motors that move with regular steps.
Journal ArticleDOI

Direct mechanical measurements of the elasticity of single DNA molecules by using magnetic beads

TL;DR: Deviations from the force curves predicted by the freely jointed chain model suggest that DNA has significant local curvature in solution, and the effect of bend-inducing cis-diamminedichloroplatinum (II) was large and supports the hypothesis of natural curvatures in DNA.
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