Abstract
Carbenes are highly reactive chemical compounds that are exploited as ligands in organometallic chemistry and are powerful organic catalysts. They were postulated to occur as transient intermediates in enzymes, yet their existence in a biological system could never be demonstrated directly. We present spectroscopic and structural data of a thiamin enzyme in a noncovalent complex with substrate, which implicate accumulation of a stable carbene as a major resonance contributor to deprotonated thiamin.
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Acknowledgements
We thank G.M. Sheldrick for providing a prerelease version of software SHELXL-2012 and discussion. We thank G. Bertrand, R. Kluger and F. Jordan for discussion and comments on the manuscript. We further thank A. Pohl and S. Lüdtke for technical assistance. This work has been supported through grants SFB 860/B07 and FOR 1296/TP3 of the Deutsche Forschungsgemeinschaft (both to K.T.).
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K.T. conceived the study; D.M. recombinantly expressed, purified and crystallized enzyme; D.M. conducted CD spectroscopy experiments; P.N. recorded the diffraction data at the European Synchrotron Radiation Facility, Grenoble, France; D.M. refined the structure; P.N. conducted final refinement rounds and estimation of estimated s.d. values using the software SHELXL-2012; D.M., P.N., R.F. and K.T. discussed the data; and K.T. wrote the paper with input from all authors.
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Meyer, D., Neumann, P., Ficner, R. et al. Observation of a stable carbene at the active site of a thiamin enzyme. Nat Chem Biol 9, 488–490 (2013). https://doi.org/10.1038/nchembio.1275
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DOI: https://doi.org/10.1038/nchembio.1275
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