Nanopores -- a Versatile Tool to Study Protein Dynamics

TL;DR

This paper reviews how nanopore technology can be used as a versatile, label-free method to study protein dynamics across a wide range of timescales, offering insights beyond traditional techniques.

Contribution

It introduces electrical nanopore detection as a powerful tool for probing protein conformational changes and interactions, highlighting its advantages and potential applications.

Findings

Nanopores provide label-free, high temporal resolution detection of protein dynamics.

Comparison shows nanopores can access timescales from microseconds to hours.

The review encourages broader adoption of nanopore techniques in protein research.

Abstract

Proteins are the active working horses in our body. These biomolecules perform all vital cellular functions from DNA replication and general biosynthesis to metabolic signaling and environmental sensing. While static 3D structures are now readily available, observing the functional cycle of proteins - involving conformational changes and interactions - remains very challenging, e.g., due to ensemble averaging. However, time-resolved information is crucial to gain a mechanistic understanding of protein function. Single-molecule techniques such as FRET and force spectroscopies provide answers but can be limited by the required labelling, a narrow time bandwidth, and more. Here, we describe electrical nanopore detection as a tool for probing protein dynamics. With a time bandwidth ranging from microseconds to hours, it covers an exceptionally wide range of timescales that is very relevant for protein function. First, we discuss the working principle of label-free nanopore experiments, various pore designs, instrumentation, and the characteristics of nanopore signals. In the second part, we review a few nanopore experiments that solved research questions in protein science, and we compare nanopores to other single-molecule techniques. We hope to make electrical nanopore sensing more accessible to the biochemical community, and to inspire new creative solutions to resolve a variety of protein dynamics - one molecule at a time.