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Title: Cleavage of a carbon–fluorine bond by an engineered cysteine dioxygenase
Cysteine dioxygenase (CDO) plays an essential role in sulfur metabolism by regulating homeostatic levels of cysteine. Human CDO contains a post-translationally generated Cys93–Tyr157 cross-linked cofactor. Here, we investigated this Cys–Tyr cross-linking by incorporating unnatural tyrosines in place of Tyr157 via a genetic method. The catalytically active variants were obtained with a thioether bond between Cys93 and the halogen-substituted Tyr157, and we determined the crystal structures of both wild-type and engineered CDO variants in the purely uncross-linked form and with a mature cofactor. Along with mass spectrometry and 19F NMR, these data indicated that the enzyme could catalyze oxidative C–F or C–Cl bond cleavage, resulting in a substantial conformational change of both Cys93 and Tyr157 during cofactor assembly. These findings provide insights into the mechanism of Cys–Tyr cofactor biogenesis and may aid the development of bioinspired aromatic carbon–halogen bond activation.  more » « less
Award ID(s):
1623856 1808637
PAR ID:
10057634
Author(s) / Creator(s):
; ; ; ; ; ; ; ;
Date Published:
Journal Name:
Nature Chemical Biology
Volume:
14
Issue:
9
ISSN:
1552-4450
Page Range / eLocation ID:
853-860
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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