TY - JOUR
T1 - YcaO domains use ATP to activate amide backbones during peptide cyclodehydrations
AU - Dunbar, Kyle L.
AU - Melby, Joel O.
AU - Mitchell, Douglas A.
N1 - Funding Information:
We are grateful to W. van der Donk for technical advice and for suggestions in the preparation of the manuscript. We thank members of the Mitchell lab and M. Marletta for critical review of this manuscript. This work was supported in part by the institutional funds provided by the University of Illinois and the US National Institutes of Health (NIH) (1R01 GM097142 to D.A.M). D.A.M. is also the recipient of the NIH Director’s New Innovator Award (DP2 OD008463). K.L.D. was supported by the NIH Training Program in the Chemistry-Biology Interface (2T32 GM070421). J.O.M. was supported by the University of Illinois Department of Chemistry Chinoree T. Kimiyo Enta Fellowship.
PY - 2012/6
Y1 - 2012/6
N2 - Thiazole/oxazole-modified microcins (TOMMs) encompass a recently defined class of ribosomally synthesized natural products with a diverse set of biological activities. Although TOMM biosynthesis has been investigated for over a decade, the mechanism of heterocycle formation by the synthetase enzymes remains poorly understood. Using substrate analogs and isotopic labeling, we demonstrate that ATP is used to directly phosphorylate the peptide amide backbone during TOMM heterocycle formation. Moreover, we present what is to our knowledge the first experimental evidence that the D-protein component of the heterocycle-forming synthetase (YcaO/domain of unknown function 181 family member), formerly annotated as a docking protein involved in complex formation and regulation, is able to perform the ATP-dependent cyclodehydration reaction in the absence of the other TOMM biosynthetic proteins. Together, these data reveal the role of ATP in the biosynthesis of azole and azoline heterocycles in ribosomal natural products and prompt a reclassification of the enzymes involved in their installation.
AB - Thiazole/oxazole-modified microcins (TOMMs) encompass a recently defined class of ribosomally synthesized natural products with a diverse set of biological activities. Although TOMM biosynthesis has been investigated for over a decade, the mechanism of heterocycle formation by the synthetase enzymes remains poorly understood. Using substrate analogs and isotopic labeling, we demonstrate that ATP is used to directly phosphorylate the peptide amide backbone during TOMM heterocycle formation. Moreover, we present what is to our knowledge the first experimental evidence that the D-protein component of the heterocycle-forming synthetase (YcaO/domain of unknown function 181 family member), formerly annotated as a docking protein involved in complex formation and regulation, is able to perform the ATP-dependent cyclodehydration reaction in the absence of the other TOMM biosynthetic proteins. Together, these data reveal the role of ATP in the biosynthesis of azole and azoline heterocycles in ribosomal natural products and prompt a reclassification of the enzymes involved in their installation.
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U2 - 10.1038/nchembio.944
DO - 10.1038/nchembio.944
M3 - Article
C2 - 22522320
AN - SCOPUS:84861308350
SN - 1552-4450
VL - 8
SP - 569
EP - 575
JO - Nature Chemical Biology
JF - Nature Chemical Biology
IS - 6
ER -