The DOI matches its Crossref record; 2 of the 2 references it cites were found in a registry or an index.
A Combined Approach for the Localization and Tandem Affinity Purification of Protein Complexes from Metazoans
Abstract
An understanding of a protein's function is greatly aided by the knowledge of its localization in vivo and identification of any interacting partners. Here, we describe a method, based on expression of a genetically encoded fusion protein, that allows for protein localization and affinity purification (LAP) in metazoans. This method makes it possible to rapidly identify transformants and to conduct live imaging of protein localization and dynamics. In addition, the same tag can be used in a modified tandem affinity purification (TAP) procedure to isolate native protein complexes of high purity. The efficacy of this purification procedure allows for the characterization of rare protein complexes that are largely inaccessible to classical biochemical purifications. The LAP strategy should be widely applicable to the characterization of protein function in cellular and developmental contexts in various metazoans.
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Cites
Global analysis of protein localization in budding yeast
Huh, W.-K., Falvo, J. V., Gerke, L. C., Carroll, A. S., Howson, R. W., Weissman, J. S., & O'Shea, E. K. (2003) · Nature · doi:10.1038/nature02026 · Cited by 4,248 · cited 2×
“Although it is possible that fusing the 26kD GFP to the N or C terminus of a protein may interfere with some aspects of its function, genome-wide studies in budding yeast have demonstrated that more than 75% of the proteins are functional with a C-terminal GFP tag (8).”
BackgroundGlobal analysis of protein expression in yeast
Ghaemmaghami, S., Huh, W.-K., Bower, K., Howson, R. W., Belle, A., Dephoure, N., O'Shea, E. K., & Weissman, J. S. (2003) · Nature · doi:10.1038/nature02046 · Cited by 3,750
Cited by
Nachury, M. V., Loktev, A. V., Zhang, Q., Westlake, C. J., Peränen, J., Merdes, A., Slusarski, D. C., Scheller, R. H., Bazan, J. F., Sheffield, V. C., et al. (2007) · Cell · doi:10.1016/j.cell.2007.03.053 · Cited by 1,384
Polo-like kinase-1 is activated by aurora A to promote checkpoint recovery
Macůrek, L., Lindqvist, A., Lim, D., Lampson, M. A., Klompmaker, R., Freire, R., Clouin, C., Taylor, S. S., Yaffe, M. B., & Medema, R. H. (2008) · Nature · doi:10.1038/nature07185 · Cited by 710
Mukhopadhyay, S., Wen, X., Chih, B., Nelson, C. D., Lane, W. S., Scales, S. J., & Jackson, P. K. (2010) · Genes & Development · doi:10.1101/gad.1966210 · Cited by 435
Quantitative proteomics combined with BAC TransgeneOmics reveals in vivo protein interactions
Hubner, N. C., Bird, A. W., Cox, J., Splettstoesser, B., Bandilla, P., Poser, I., Hyman, A., & Mann, M. (2010) · Journal of Cell Biology · doi:10.1083/jcb.200911091 · Cited by 440
An efficient tandem affinity purification procedure for interaction proteomics in mammalian cells
Bürckstümmer, T., Bennett, K. L., Preradovic, A., Schütze, G., Hantschel, O., Superti-Furga, G., & Bauch, A. (2006) · Nature Methods · doi:10.1038/nmeth968 · Cited by 391
Metadata
- Authors
- Cheeseman, I. M., & Desai, A.
- Venue
- Science's STKE
- Published
- 2005-01-11
- Volume
- 2005
- Issue
- 266
- Pages
- pl1-pl1
- ISSN
- 1525-8882 (ISSN-L)
- OpenAlex
- W2108025706
- Cited by
- 254
- References
- 2
- FWCI
- 3.83
- Type
- journal article
- Language
- en
- Open access
- closed
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