Proteomic navigation using proximity-labeling.
Loading...
Embargo End Date
ICR Authors
Authors
Gentzel, M
Pardo, M
Subramaniam, S
Stewart, AF
Choudhary, JS
Pardo, M
Subramaniam, S
Stewart, AF
Choudhary, JS
Document Type
Journal Article
Date
2019-07-15
Date Accepted
2019-03-29
Abstract
The identification of bona fide protein-protein interactions and the mapping of proteomes was greatly enhanced by protein tagging for generic affinity purification methods and analysis by mass spectrometry (AP-MS). The high quality of AP-MS data permitted the development of proteomic navigation by sequential tagging of identified interactions. However AP-MS is laborious and limited to relatively high affinity protein-protein interactions. Proximity labeling, first with the biotin ligase BirA, termed BioID, and then with ascorbate peroxidase, termed APEX, permits a greater reach into the proteome than AP-MS enabling both the identification of a wider field and weaker protein-protein interactions. This additional reach comes with the need for stringent controls. Proximity labeling also permits experiments in living cells allowing spatiotemporal investigations of the proteome. Here we discuss proximity labeling with accompanying methodological descriptions for E. coli and mammalian cells.
Citation
Methods (San Diego, Calif.), 2019, 164-165 pp. 67 - 72
Source Title
Publisher
ACADEMIC PRESS INC ELSEVIER SCIENCE
ISSN
1046-2023
eISSN
1095-9130
Collections
Research Team
Functional Proteomics Group