Methods for introducing bioorthogonal functionalities into proteins have become central to protein engineering efforts. Here we describe a method for the site-specific introduction of aldehyde groups into recombinant proteins using the 6-amino-acid consensus sequence recognized by the formylglycine-generating enzyme. This genetically encoded 'aldehyde tag' is no larger than a His6 tag and can be exploited for numerous protein labeling applications.
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We thank M. Francis and J. Rush for helpful discussions and D. King and A. Falick for MS expertise. I.S.C. was supported by a postdoctoral fellowship from the US National Institutes of Health. B.L.C. was supported by a predoctoral fellowship from the US National Science Foundation. This work was supported by a grant from the US National Institutes of Health to C.R.B. (GM59907).
The authors declare no competing financial interests.
Mass spectra confirming the presence of fGly in a tryptic peptide from ald13-Stf0. (PDF 222 kb)
Quantitation of the conversion of cysteine to formylglycine using mass spectrometry. (PDF 156 kb)
Quantitation of the conversion of cysteine to formylglycine using Alexa Fluor 647 C5-aminooxyacetamide labeling. (PDF 251 kb)
PEGylation of ald6-MBP with 2 kDa and 5 kDa aminooxy-PEG. (PDF 184 kb)
Oligonucleotides used in this study. (PDF 274 kb)
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Carrico, I., Carlson, B. & Bertozzi, C. Introducing genetically encoded aldehydes into proteins. Nat Chem Biol 3, 321–322 (2007). https://doi.org/10.1038/nchembio878
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