The DOI matches its Crossref record; 1 of the 1 references it cites were found in a registry or an index.
Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli
Open accessAbstract
Escherichia coli is one of the most widely used vehicles to overexpress membrane proteins (MPs). Currently, it is not possible to predict if an overexpressed MP will end up in the cytoplasmic membrane or in inclusion bodies. Overexpression of MPs in the cytoplasmic membrane is strongly favoured to overexpression in inclusion bodies, since it is relatively easy to isolate MPs from membranes and usually impossible to isolate them from inclusion bodies. Here we show that green fluorescent protein (GFP), when fused to an overexpressed MP, can be used as an indicator to monitor membrane insertion versus inclusion body formation of overexpressed MPs in E. coli. Furthermore, we show that an overexpressed MP can be recovered from a MP–GFP fusion using a site specific protease. This makes GFP an excellent tool for large‐scale MP target selection in structural genomics projects.
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Cites
Membrane Topology of the 60-kDa Oxa1p Homologue fromEscherichia coli
Sääf, A., Monné, M., de Gier, J.-W., & von Heijne, G. (1998) · Journal of Biological Chemistry · doi:10.1074/jbc.273.46.30415 · Cited by 89
Cited by
Global Topology Analysis of the Escherichia coli Inner Membrane Proteome
Daley, D. O., Rapp, M., Granseth, E., Melén, K., Drew, D., & von Heijne, G. (2005) · Science · doi:10.1126/science.1109730 · Cited by 490
Self-citationOptimization of membrane protein overexpression and purification using GFP fusions
Drew, D., Lerch, M., Kunji, E., Slotboom, D.-J., & de Gier, J.-W. (2006) · Nature Methods · doi:10.1038/nmeth0406-303 · Cited by 339
Self-citationCryo-EM structure and resistance landscape of M. tuberculosis MmpL3: An emergent therapeutic target
Adams, O., Deme, J. C., Parker, J. L., Fowler, P. W., Lea, S. M., & Newstead, S. (2021) · Structure · doi:10.1016/j.str.2021.06.013 · Cited by 63
Oleinikov, A. V., Rossnagle, E., Francis, S., Mutabingwa, T. K., Fried, M., & Duffy, P. E. (2007) · The Journal of Infectious Diseases · doi:10.1086/518513 · Cited by 59
Metadata
- Authors
- Drew, D. E., von Heijne, G., Nordlund, P., & de Gier, J.-W. L.
- Venue
- FEBS Letters
- Publisher
- Wiley
- Published
- 2001-10-15
- Volume
- 507
- Issue
- 2
- Pages
- 220-224
- ISSN
- 0014-5793 (ISSN-L)
- 1873-3468
- OpenAlex
- W2000896683
- Cited by
- 236
- References
- 1
- FWCI
- 3.50
- Type
- journal article
- Language
- en
- Open access
- bronze
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- Drew, D. E., von Heijne, G., Nordlund, P., & de Gier, J.-W. L. (2001). Green fluorescent protein as an indicator to monitor membrane protein overexpression in Escherichia coli. FEBS Letters. https://doi.org/10.1016/s0014-5793(01)02980-5
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Contains information from the Semantic Scholar Academic Graph Datasets, made available under the ODC Attribution License (ODC-BY 1.0).
Semantic Scholar’s data is described in: Kinney, R., et al. (2023). The Semantic Scholar Open Data Platform. arXiv:2301.10140.
