Single-Molecule Mechanics in Ligand Concentration Gradient

Kretzer, Balázs; Kiss, Bálint; Tordai, Hedvig; Csík, Gabriella; Herényi, Levente; Kellermayer, Miklós

doi:10.3390/mi11020212

Open AccessArticle

Single-Molecule Mechanics in Ligand Concentration Gradient

by

Balázs Kretzer

,

Bálint Kiss

,

Hedvig Tordai

,

Gabriella Csík

,

Levente Herényi

and

Miklós Kellermayer

^*

Department of Biophysics and Radiation Biology, Semmelweis University, 1094 Budapest, Hungary

^*

Author to whom correspondence should be addressed.

Micromachines 2020, 11(2), 212; https://doi.org/10.3390/mi11020212

Submission received: 28 December 2019 / Revised: 9 February 2020 / Accepted: 14 February 2020 / Published: 19 February 2020

(This article belongs to the Special Issue Optical Trapping and Manipulation: From Fundamentals to Applications)

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Abstract

Single-molecule experiments provide unique insights into the mechanisms of biomolecular phenomena. However, because varying the concentration of a solute usually requires the exchange of the entire solution around the molecule, ligand-concentration-dependent measurements on the same molecule pose a challenge. In the present work we exploited the fact that a diffusion-dependent concentration gradient arises in a laminar-flow microfluidic device, which may be utilized for controlling the concentration of the ligand that the mechanically manipulated single molecule is exposed to. We tested this experimental approach by exposing a λ-phage dsDNA molecule, held with a double-trap optical tweezers instrument, to diffusionally-controlled concentrations of SYTOX Orange (SxO) and tetrakis(4-N-methyl)pyridyl-porphyrin (TMPYP). We demonstrate that the experimental design allows access to transient-kinetic, equilibrium and ligand-concentration-dependent mechanical experiments on the very same single molecule.

Keywords: optical tweezers; concentration gradient; force spectroscopy; diffusion; microfluidics; fluorescence

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MDPI and ACS Style

Kretzer, B.; Kiss, B.; Tordai, H.; Csík, G.; Herényi, L.; Kellermayer, M. Single-Molecule Mechanics in Ligand Concentration Gradient. Micromachines 2020, 11, 212. https://doi.org/10.3390/mi11020212

AMA Style

Kretzer B, Kiss B, Tordai H, Csík G, Herényi L, Kellermayer M. Single-Molecule Mechanics in Ligand Concentration Gradient. Micromachines. 2020; 11(2):212. https://doi.org/10.3390/mi11020212

Chicago/Turabian Style

Kretzer, Balázs, Bálint Kiss, Hedvig Tordai, Gabriella Csík, Levente Herényi, and Miklós Kellermayer. 2020. "Single-Molecule Mechanics in Ligand Concentration Gradient" Micromachines 11, no. 2: 212. https://doi.org/10.3390/mi11020212

APA Style

Kretzer, B., Kiss, B., Tordai, H., Csík, G., Herényi, L., & Kellermayer, M. (2020). Single-Molecule Mechanics in Ligand Concentration Gradient. Micromachines, 11(2), 212. https://doi.org/10.3390/mi11020212

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Single-Molecule Mechanics in Ligand Concentration Gradient

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