The DOI matches its Crossref record; 5 of the 5 references it cites were found in a registry or an index.
A general protocol for the generation of Nanobodies for structural biology
Open accessAbstract
There is growing interest in using antibodies as auxiliary tools to crystallize proteins. Here we describe a general protocol for the generation of Nanobodies to be used as crystallization chaperones for the structural investigation of diverse conformational states of flexible (membrane) proteins and complexes thereof. Our technology has a competitive advantage over other recombinant crystallization chaperones in that we fully exploit the natural humoral response against native antigens. Accordingly, we provide detailed protocols for the immunization with native proteins and for the selection by phage display of in vivo–matured Nanobodies that bind conformational epitopes of functional proteins. Three representative examples illustrate that the outlined procedures are robust, making it possible to solve by Nanobody-assisted X-ray crystallography in a time span of 6–12 months.
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Naturally occurring antibodies devoid of light chains
Hamers-Casterman, C., Atarhouch, T., Muyldermans, S., Robinson, G., Hammers, C., Songa, E. B., Bendahman, N., & Hammers, R. (1993) · Nature · doi:10.1038/363446a0 · Cited by 3,241
Self-citationCrystal structure of the β2 adrenergic receptor–Gs protein complex
Rasmussen, S. G. F., DeVree, B. T., Zou, Y., Kruse, A. C., Chung, K. Y., Kobilka, T. S., Thian, F. S., Chae, P. S., Pardon, E., Calinski, D., et al. (2011) · Nature · doi:10.1038/nature10361 · Cited by 3,009
Self-citationNanobodies: natural single-domain antibodies
Muyldermans, S. (2013) · Annual Review of Biochemistry · doi:10.1146/annurev-biochem-063011-092449 · Cited by 2,304
Self-citationConformational biosensors reveal GPCR signalling from endosomes
Irannejad, R., Tomshine, J. C., Tomshine, J. R., Chevalier, M., Mahoney, J. P., Steyaert, J., Rasmussen, S. G. F., Sunahara, R. K., El-Samad, H., Huang, B., et al. (2013) · Nature · doi:10.1038/nature12000 · Cited by 819
Self-citationNanobody stabilization of G protein-coupled receptor conformational states
Steyaert, J., & Kobilka, B. K. (2011) · Current Opinion in Structural Biology · doi:10.1016/j.sbi.2011.06.011 · Cited by 234
Cited by
Common activation mechanism of class A GPCRs
Zhou, Q., Yang, D., Wu, M., Guo, Y., Guo, W., Zhong, L., Cai, X., Dai, A., Jang, W., Shakhnovich, E. I., et al. (2019) · eLife · doi:10.7554/elife.50279 · Cited by 632
Structure of the Nanobody-Stabilized Active State of the Kappa Opioid Receptor
Che, T., Majumdar, S., Zaidi, S. A., Ondachi, P., McCorvy, J. D., Wang, S., Mosier, P. D., Uprety, R., Vardy, E., Krumm, B. E., et al. (2018) · Cell · doi:10.1016/j.cell.2017.12.011 · Cited by 365
Self-citationStructural insights into the activation of metabotropic glutamate receptors
Koehl, A., Hu, H., Feng, D., Sun, B., Zhang, Y., Robertson, M. J., Chu, M., Kobilka, T. S., Laeremans, T., Steyaert, J., et al. (2019) · Nature · doi:10.1038/s41586-019-0881-4 · Cited by 325
Self-citationThe Molecular Mechanism of Transport by the Mitochondrial ADP/ATP Carrier
Ruprecht, J. J., King, M. S., Zögg, T., Aleksandrova, A. A., Pardon, E., Crichton, P. G., Steyaert, J., & Kunji, E. R. S. (2019) · Cell · doi:10.1016/j.cell.2018.11.025 · Cited by 319
Self-citationNanobodies to Study G Protein–Coupled Receptor Structure and Function
Manglik, A., Kobilka, B. K., & Steyaert, J. (2017) · Annual Review of Pharmacology and Toxicology · doi:10.1146/annurev-pharmtox-010716-104710 · Cited by 257
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Metadata
- Authors
- Pardon, E., Laeremans, T., Triest, S., Rasmussen, S. G. F., Wohlkönig, A., Ruf, A., Muyldermans, S., Hol, W. G. J., Kobilka, B. K., & Steyaert, J.
- Venue
- Nature Protocols
- Published
- 2014-02-27
- Volume
- 9
- Issue
- 3
- Pages
- 674-693
- ISSN
- 1750-2799 (ISSN-L)
- 1754-2189
- OpenAlex
- W1998292458
- Cited by
- 790
- References
- 5
- FWCI
- 18.61
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Pardon, E., Laeremans, T., Triest, S., Rasmussen, S. G. F., Wohlkönig, A., Ruf, A., Muyldermans, S., Hol, W. G. J., Kobilka, B. K., & Steyaert, J. (2014). A general protocol for the generation of Nanobodies for structural biology. Nature Protocols. https://doi.org/10.1038/nprot.2014.039
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
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