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Genetically encoded protein sulfation in mammalian cells


Tyrosine sulfation is an important post-translational modification found in higher eukaryotes. Here we report an engineered tyrosyl-tRNA synthetase/tRNA pair that co-translationally incorporates O-sulfotyrosine in response to UAG codons in Escherichia coli and mammalian cells. This platform enables recombinant expression of eukaryotic proteins homogeneously sulfated at chosen sites, which was demonstrated by expressing human heparin cofactor II in mammalian cells in different states of sulfation.

Fig. 1: Genetically encoding sTyr.
Fig. 2: Expression and biochemical analysis of precisely sulfated HCII.

Data availability

Data associated with this work are available upon request from the corresponding author.


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We thank D.M. Monroe III (UNC) for helpful discussions on the HCII kinetic assay. This work was supported by the National Institutes of Health (R01GM124319 and R01GM126220 to A.C.; 1R01GM118431 and 1R01GM117004 to E.W.).

Author information




A.C. and J.S.I. designed the experiments. J.S.I. conducted all experiments. C.M.H. and C.L. assisted with cloning and protein expression. J.C.P. performed the MS analyses of HCII and E.W. supervised. A.C. and J.S.I. prepared the manuscript.

Corresponding author

Correspondence to Abhishek Chatterjee.

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The authors declare no competing interests.

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Supplementary Information

Supplementary Figs. 1–11, Table 1 and Note.

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Italia, J.S., Peeler, J.C., Hillenbrand, C.M. et al. Genetically encoded protein sulfation in mammalian cells. Nat Chem Biol 16, 379–382 (2020).

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