Abstract
Integral membrane proteins are important drug targets that are critical in supporting many biological processes. Despite that, the study of their structure–function relationships remains a major goal in structural biology, yet progress has been hampered by inherent challenges in the production for stable and homogeneous protein samples. Dynamic light scattering provides a straightforward probe of protein quality in solution, particularly in relation to stability and aggregation. However, the necessity to use large amounts of sample and the low-throughput nature of the analysis remain as major bottlenecks of the technique.
Here, we present a protocol for dynamic light scattering measurements that are executed in a fully automated fashion for low-volume samples, in situ. The protocol offers a generic pre-screening method for downstream structural studies of biomolecules using higher-resolution approaches such as X-ray crystallography, electron microscopy, small-angle X-ray scattering, and NMR .
Key words
- Dynamic light scattering
- Membrane proteins and biophysics
- Biomembrane interfaces
- Biomolecules
- Protein solubility and aggregation
- Crystal nucleation
- Monodispersity
- Hydrodynamic radius
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Acknowledgments
We acknowledge funding from the United Kingdom’s Department of Business, Energy and Industrial Strategy (BEIS).
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Kwan, T.O.C., Reis, R., Moraes, I. (2021). In Situ Measurements of Polypeptide Samples by Dynamic Light Scattering: Membrane Proteins, a Case Study. In: Ryadnov, M. (eds) Polypeptide Materials. Methods in Molecular Biology, vol 2208. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0928-6_13
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DOI: https://doi.org/10.1007/978-1-0716-0928-6_13
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