Abstract
Attempts to elucidate the roles of carbohydrate-associated structures in biology have led to the distinct field of glycobiology research. The focus of this field has been in understanding the evolution, biosynthesis and interactions of glycans, both individually and as components of larger biomolecules. However, as most approaches for studying glycans (including mass spectrometry and various binding assays) use ensemble measurements, they lack the precision required to uncover the discrete roles of glycoconjugates, which are often heterogeneous, in biomolecular processes. Single-molecule techniques can examine individual events within challenging mixtures, and they are beginning to be applied to glycobiology. For example, single-molecule force spectroscopy (SMFS) by atomic force microscopy (AFM) has enabled the molecular interactions of sugars to be studied, single-molecule fluorescence microscopy and spectroscopy have led to insight into the role of sugars in biological processes and nanopores have revealed interactions between polysaccharides and their transporters. Thus, single-molecule technology is becoming a valuable tool in glycoscience.
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The authors thank S. Faulkner, M. Bilyard and K. Wals for proofreading and useful comments.
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Nature Reviews Chemistry thanks Xing Chen, Isabelle Compagnon, Lara Mahal and Bingqian Xu for their contribution to the peer review of this work.
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Glossary
- Force–distance cycles
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The repeated measurement of force as a function of distance in atomic force microscopy.
- Total internal reflection fluorescence
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(TIRF). Microscopy examining a thin region of a specimen (~100 nm) that uses internal reflection at a surface–sample interface for the selective excitation of surface-associated fluorophores, thereby removing contributions from the background.
- Stochastic optical reconstruction microscopy
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(STORM). Dilute fluorophores are switched on and off through switching or bleaching, and the ‘centres’ of their location are ascertained using point-spread of photon emission based on a widefield image. After several rounds, a ‘super-resolution’ image is assembled by plotting the measured positions of these centres of the fluorescent probes.
- Photoactivated localization microscopy
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(PALM). Similar to STORM but has typically used photoactivation and then bleaching to switch fluorophores on and off. It is more often associated with the use of protein fluorophores such as photoactivatable fluorescent proteins.
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Lakshminarayanan, A., Richard, M. & Davis, B.G. Studying glycobiology at the single-molecule level. Nat Rev Chem 2, 148–159 (2018). https://doi.org/10.1038/s41570-018-0019-5
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DOI: https://doi.org/10.1038/s41570-018-0019-5
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