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Single-molecule approach to immunoprecipitated protein complexes: Insights into miRNA uridylation
Cited 63 time in
Web of Science
Cited 66 time in Scopus
- Authors
- Issue Date
- 2011-07
- Publisher
- Nature Publishing Group
- Citation
- EMBO Reports, Vol.12 No.7, pp.690-696
- Abstract
- Single-molecule techniques have been used for only a subset of biological problems because of difficulties in studying proteins that require cofactors or post-translational modifications. Here, we present a new method integrating single-molecule fluorescence microscopy and immunopurification to study protein complexes. We used this method to investigate Lin28-mediated microRNA uridylation by TUT4 (terminal uridylyl transferase 4, polyU polymerase), which regulates let-7 microRNA biogenesis. Our real-time analysis of the uridylation by the TUT4 immunoprecipitates suggests that Lin28 functions as a processivity factor of TUT4. Our new technique, SIMPlex (single-molecule approach to immunoprecipitated protein complexes), provides a universal tool to analyse complex proteins at the single-molecule level.
- ISSN
- 1469-221X
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