Rapid Microfluidic Dilution for Single-Molecule Spectroscopy of Low-Affinity Biomolecular Complexes

Angewandte Chemie
Niels ZijlstraBenjamin Schuler

Abstract

To enable the investigation of low-affinity biomolecular complexes with confocal single-molecule spectroscopy, we have developed a microfluidic device that allows a concentrated sample to be diluted by up to five orders of magnitude within milliseconds, at the physical limit dictated by diffusion. We demonstrate the capabilities of the device by studying the dissociation kinetics and structural properties of low-affinity protein complexes using single-molecule two-color and three-color Förster resonance energy transfer (FRET). We show that the versatility of the device makes it suitable for studying complexes with dissociation constants from low nanomolar up to 10 μm, thus covering a wide range of biomolecular interactions. The design and precise fabrication of the devices ensure simple yet reliable operation and high reproducibility of the results.

References

Jul 18, 2002·Accounts of Chemical Research·J Cooper McDonald, George M Whitesides
Apr 17, 2008·Annual Review of Biochemistry·Chirlmin JooTaekjip Ha
Feb 7, 2009·Chemical Reviews·G SchreiberH-X Zhou
Jun 3, 2009·The Review of Scientific Instruments·Shawn H PfeilEverett A Lipman
Jul 6, 2010·Molecular BioSystems·Yann Gambin, Ashok A Deniz
Jul 10, 2010·Proceedings of the National Academy of Sciences of the United States of America·Magnus KjaergaardFlemming M Poulsen
Nov 15, 2011·Nature Methods·Roger B AltmanScott C Blanchard
Jan 15, 2013·Current Opinion in Structural Biology·Benjamin Schuler, Hagen Hofmann
Jun 21, 2013·Analytical Chemistry·Mathew H HorrocksDavid Klenerman
Aug 29, 2014·Proceedings of the National Academy of Sciences of the United States of America·Ruth KellnerBenjamin Schuler
Feb 6, 2015·Nature Communications·Stephan BenkeBenjamin Schuler
Feb 7, 2016·The Journal of Biological Chemistry·Sarah L ShammasJane Clarke
Feb 23, 2017·Proceedings of the National Academy of Sciences of the United States of America·Andrea SorannoBenjamin Schuler

❮ Previous
Next ❯

Citations

Mar 14, 2020·Angewandte Chemie·Fabian SoltermannWeston B Struwe
Aug 22, 2018·Nature Communications·Franziska ZoselBenjamin Schuler
Apr 9, 2020·Biophysics Reviews·William E ArterTuomas P J Knowles
Jan 30, 2021·Chem·Anna OlerinyovaPhilipp Kukura

❮ Previous
Next ❯

Related Concepts

Trending Feeds

COVID-19

Coronaviruses encompass a large family of viruses that cause the common cold as well as more serious diseases, such as the ongoing outbreak of coronavirus disease 2019 (COVID-19; formally known as 2019-nCoV). Coronaviruses can spread from animals to humans; symptoms include fever, cough, shortness of breath, and breathing difficulties; in more severe cases, infection can lead to death. This feed covers recent research on COVID-19.

Blastomycosis

Blastomycosis fungal infections spread through inhaling Blastomyces dermatitidis spores. Discover the latest research on blastomycosis fungal infections here.

Nuclear Pore Complex in ALS/FTD

Alterations in nucleocytoplasmic transport, controlled by the nuclear pore complex, may be involved in the pathomechanism underlying multiple neurodegenerative diseases including Amyotrophic Lateral Sclerosis and Frontotemporal Dementia. Here is the latest research on the nuclear pore complex in ALS and FTD.

Applications of Molecular Barcoding

The concept of molecular barcoding is that each original DNA or RNA molecule is attached to a unique sequence barcode. Sequence reads having different barcodes represent different original molecules, while sequence reads having the same barcode are results of PCR duplication from one original molecule. Discover the latest research on molecular barcoding here.

Chronic Fatigue Syndrome

Chronic fatigue syndrome is a disease characterized by unexplained disabling fatigue; the pathology of which is incompletely understood. Discover the latest research on chronic fatigue syndrome here.

Evolution of Pluripotency

Pluripotency refers to the ability of a cell to develop into three primary germ cell layers of the embryo. This feed focuses on the mechanisms that underlie the evolution of pluripotency. Here is the latest research.

Position Effect Variegation

Position Effect Variagation occurs when a gene is inactivated due to its positioning near heterochromatic regions within a chromosome. Discover the latest research on Position Effect Variagation here.

STING Receptor Agonists

Stimulator of IFN genes (STING) are a group of transmembrane proteins that are involved in the induction of type I interferon that is important in the innate immune response. The stimulation of STING has been an active area of research in the treatment of cancer and infectious diseases. Here is the latest research on STING receptor agonists.

Microbicide

Microbicides are products that can be applied to vaginal or rectal mucosal surfaces with the goal of preventing, or at least significantly reducing, the transmission of sexually transmitted infections. Here is the latest research on microbicides.