Projects per year
Abstract
Serine phosphorylation is a key post-translational modification that regulates diverse biological processes. Powerful analytical methods have identified thousands of phosphorylation sites, but many of their functions remain to be deciphered. A key to understanding the function of protein phosphorylation is access to phosphorylated proteins, but this is often challenging or impossible. Here we evolve an orthogonal aminoacyl-tRNA synthetase/tRNA<inf>CUA</inf> pair that directs the efficient incorporation of phosphoserine (pSer (1)) into recombinant proteins in Escherichia coli. Moreover, combining the orthogonal pair with a metabolically engineered E. coli enables the site-specific incorporation of a nonhydrolyzable analog of pSer. Our approach enables quantitative decoding of the amber stop codon as pSer, and we purify, with yields of several milligrams per liter of culture, proteins bearing biologically relevant phosphorylations that were previously challenging or impossible to access - including phosphorylated ubiquitin and the kinase Nek7, which is synthetically activated by a genetically encoded phosphorylation in its activation loop.
Original language | English |
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Pages (from-to) | 496-503 |
Number of pages | 8 |
Journal | Nature Chemical Biology |
Volume | 11 |
Issue number | 7 |
DOIs | |
Publication status | Published - Jul 2015 |
ASJC Scopus subject areas
- Cell Biology
- Molecular Biology
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Dive into the research topics of 'Efficient genetic encoding of phosphoserine and its nonhydrolyzable analog'. Together they form a unique fingerprint.Projects
- 3 Finished
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Structural Analysis of the Parkinson's Associated Kinase PINK1
Muqit, M. (Investigator) & van Aalten, D. (Investigator)
1/11/15 → 31/10/16
Project: Research
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Investigation of the Regulation of Rab GTPases by PINK1 (PhD Studentship)
Muqit, M. (Investigator)
7/09/15 → 6/09/18
Project: Research
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Biochemical Analysis of the PINK-1 Parkin Signalling Pathway in Parkinson's Disease (Senior Clinical Fellowship)
Muqit, M. (Investigator)
1/07/13 → 31/12/23
Project: Research
Student theses
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Regulation of the Parkin E3 ligase by PINK1-dependent phosphorylation
Kazlauskaite, A. (Author), Muqit, M. (Supervisor) & Alessi, D. (Supervisor), 2015Student thesis: Doctoral Thesis › Doctor of Philosophy
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