Protein neighbors and proximity proteomics
Rees, Johanna Susan
Lilley, Kathryn Susan
Molecular & Cellular Proteomics
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Rees, J. S., Li, X., Perrett, S., Lilley, K. S., & Jackson, A. P. (2015). Protein neighbors and proximity proteomics. Molecular & Cellular Proteomics, 14 2848-2856. https://doi.org/10.1074/mcp.R115.052902
Within cells, proteins can co-assemble into functionally integrated and spatially restricted multicomponent complexes. Often, the affinities between individual proteins are relatively weak, and proteins within such clusters may interact only indirectly with many of their other protein neighbors. This makes proteomic characterization difficult using methods such as immunoprecipitation or cross-linking. Recently, several groups have described the use of enzyme-catalyzed proximity labeling reagents that covalently tag the neighbors of a targeted protein with a small molecule such as fluorescein or biotin. The modified proteins can then be isolated by standard pulldown methods and identified by mass spectrometry. Here we will describe the techniques as well as their similarities and differences. We discuss their applications both to study protein assemblies and to provide a new way for characterizing organelle proteomes. We stress the importance of proteomic quantitation and independent target validation in such experiments. Furthermore, we suggest that there are biophysical and cell-biological principles that dictate the appropriateness of enzyme-catalyzed proximity labeling methods to address particular biological questions of interest.
This work was supported by Biotechnology and Biological Sciences Research Council (UK) Grant BB/J021091/1. XWL & SP supported by Chinese Ministry of Science and Technology 973 Program Grants 2012CB911000 and 2013CB910700 and National Natural Science Foundation of China Grant 31110103914.
External DOI: https://doi.org/10.1074/mcp.R115.052902
This record's URL: https://www.repository.cam.ac.uk/handle/1810/253300
Attribution 2.0 UK: England & Wales
Licence URL: http://creativecommons.org/licenses/by/2.0/uk/
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