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13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions

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13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions. / Lohans, Christopher T.; Wang, David Y.; Jorgensen, Christian; Cahill, Samuel T.; Clifton, Ian J.; McDonough, Michael A.; Oswin, Henry P.; Spencer, James; Domene Nunez, Carmen; Claridge, Timothy D.W.; Brem, Jürgen; Schofield, Christopher J.

In: ORGANIC AND BIOMOLECULAR CHEMISTRY, Vol. 15, No. 28, 26.06.2017, p. 6024-6032.

Research output: Contribution to journalArticle

Harvard

Lohans, CT, Wang, DY, Jorgensen, C, Cahill, ST, Clifton, IJ, McDonough, MA, Oswin, HP, Spencer, J, Domene Nunez, C, Claridge, TDW, Brem, J & Schofield, CJ 2017, '13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions', ORGANIC AND BIOMOLECULAR CHEMISTRY, vol. 15, no. 28, pp. 6024-6032. https://doi.org/10.1039/c7ob01514c

APA

Lohans, C. T., Wang, D. Y., Jorgensen, C., Cahill, S. T., Clifton, I. J., McDonough, M. A., Oswin, H. P., Spencer, J., Domene Nunez, C., Claridge, T. D. W., Brem, J., & Schofield, C. J. (2017). 13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions. ORGANIC AND BIOMOLECULAR CHEMISTRY, 15(28), 6024-6032. https://doi.org/10.1039/c7ob01514c

Vancouver

Lohans CT, Wang DY, Jorgensen C, Cahill ST, Clifton IJ, McDonough MA et al. 13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions. ORGANIC AND BIOMOLECULAR CHEMISTRY. 2017 Jun 26;15(28):6024-6032. https://doi.org/10.1039/c7ob01514c

Author

Lohans, Christopher T. ; Wang, David Y. ; Jorgensen, Christian ; Cahill, Samuel T. ; Clifton, Ian J. ; McDonough, Michael A. ; Oswin, Henry P. ; Spencer, James ; Domene Nunez, Carmen ; Claridge, Timothy D.W. ; Brem, Jürgen ; Schofield, Christopher J. / 13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions. In: ORGANIC AND BIOMOLECULAR CHEMISTRY. 2017 ; Vol. 15, No. 28. pp. 6024-6032.

Bibtex Download

@article{b72709e8202c4c5c98d06e7847a4b025,
title = "13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions",
abstract = "The class D (OXA) serine β-lactamases are a major cause of resistance to β-lactam antibiotics. The class D enzymes are unique amongst β-lactamases because they have a carbamylated lysine that acts as a general acid/base in catalysis. Previous crystallographic studies led to the proposal that β-lactamase inhibitor avibactam targets OXA enzymes in part by promoting decarbamylation. Similarly, halide ions are proposed to inhibit OXA enzymes via decarbamylation. NMR analyses, in which the carbamylated lysines of OXA-10, -23 and -48 were 13C-labelled, indicate that reaction with avibactam does not ablate lysine carbamylation in solution. While halide ions did not decarbamylate the 13C-labelled OXA enzymes in the absence of substrate or inhibitor, avibactam-treated OXA enzymes were susceptible to decarbamylation mediated by halide ions, suggesting halide ions may inhibit OXA enzymes by promoting decarbamylation of acyl-enzyme complex. Crystal structures of the OXA-10 avibactam complex were obtained with bromide, iodide, and sodium ions bound between Trp-154 and Lys-70. Structures were also obtained wherein bromide and iodide ions occupy the position expected for the 'hydrolytic water' molecule. In contrast with some solution studies, Lys-70 was decarbamylated in these structures. These results reveal clear differences between crystallographic and solution studies on the interaction of class D β-lactamases with avibactam and halides, and demonstrate the utility of 13C-NMR for studying lysine carbamylation in solution.",
author = "Lohans, {Christopher T.} and Wang, {David Y.} and Christian Jorgensen and Cahill, {Samuel T.} and Clifton, {Ian J.} and McDonough, {Michael A.} and Oswin, {Henry P.} and James Spencer and {Domene Nunez}, Carmen and Claridge, {Timothy D.W.} and J{\"u}rgen Brem and Schofield, {Christopher J.}",
year = "2017",
month = jun,
day = "26",
doi = "10.1039/c7ob01514c",
language = "English",
volume = "15",
pages = "6024--6032",
journal = "ORGANIC AND BIOMOLECULAR CHEMISTRY",
issn = "1477-0520",
publisher = "Royal Society of Chemistry",
number = "28",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - 13C-Carbamylation as a mechanistic probe for the inhibition of class D β-lactamases by avibactam and halide ions

AU - Lohans, Christopher T.

AU - Wang, David Y.

AU - Jorgensen, Christian

AU - Cahill, Samuel T.

AU - Clifton, Ian J.

AU - McDonough, Michael A.

AU - Oswin, Henry P.

AU - Spencer, James

AU - Domene Nunez, Carmen

AU - Claridge, Timothy D.W.

AU - Brem, Jürgen

AU - Schofield, Christopher J.

PY - 2017/6/26

Y1 - 2017/6/26

N2 - The class D (OXA) serine β-lactamases are a major cause of resistance to β-lactam antibiotics. The class D enzymes are unique amongst β-lactamases because they have a carbamylated lysine that acts as a general acid/base in catalysis. Previous crystallographic studies led to the proposal that β-lactamase inhibitor avibactam targets OXA enzymes in part by promoting decarbamylation. Similarly, halide ions are proposed to inhibit OXA enzymes via decarbamylation. NMR analyses, in which the carbamylated lysines of OXA-10, -23 and -48 were 13C-labelled, indicate that reaction with avibactam does not ablate lysine carbamylation in solution. While halide ions did not decarbamylate the 13C-labelled OXA enzymes in the absence of substrate or inhibitor, avibactam-treated OXA enzymes were susceptible to decarbamylation mediated by halide ions, suggesting halide ions may inhibit OXA enzymes by promoting decarbamylation of acyl-enzyme complex. Crystal structures of the OXA-10 avibactam complex were obtained with bromide, iodide, and sodium ions bound between Trp-154 and Lys-70. Structures were also obtained wherein bromide and iodide ions occupy the position expected for the 'hydrolytic water' molecule. In contrast with some solution studies, Lys-70 was decarbamylated in these structures. These results reveal clear differences between crystallographic and solution studies on the interaction of class D β-lactamases with avibactam and halides, and demonstrate the utility of 13C-NMR for studying lysine carbamylation in solution.

AB - The class D (OXA) serine β-lactamases are a major cause of resistance to β-lactam antibiotics. The class D enzymes are unique amongst β-lactamases because they have a carbamylated lysine that acts as a general acid/base in catalysis. Previous crystallographic studies led to the proposal that β-lactamase inhibitor avibactam targets OXA enzymes in part by promoting decarbamylation. Similarly, halide ions are proposed to inhibit OXA enzymes via decarbamylation. NMR analyses, in which the carbamylated lysines of OXA-10, -23 and -48 were 13C-labelled, indicate that reaction with avibactam does not ablate lysine carbamylation in solution. While halide ions did not decarbamylate the 13C-labelled OXA enzymes in the absence of substrate or inhibitor, avibactam-treated OXA enzymes were susceptible to decarbamylation mediated by halide ions, suggesting halide ions may inhibit OXA enzymes by promoting decarbamylation of acyl-enzyme complex. Crystal structures of the OXA-10 avibactam complex were obtained with bromide, iodide, and sodium ions bound between Trp-154 and Lys-70. Structures were also obtained wherein bromide and iodide ions occupy the position expected for the 'hydrolytic water' molecule. In contrast with some solution studies, Lys-70 was decarbamylated in these structures. These results reveal clear differences between crystallographic and solution studies on the interaction of class D β-lactamases with avibactam and halides, and demonstrate the utility of 13C-NMR for studying lysine carbamylation in solution.

UR - http://www.scopus.com/inward/record.url?scp=85025100632&partnerID=8YFLogxK

U2 - 10.1039/c7ob01514c

DO - 10.1039/c7ob01514c

M3 - Article

AN - SCOPUS:85025100632

VL - 15

SP - 6024

EP - 6032

JO - ORGANIC AND BIOMOLECULAR CHEMISTRY

JF - ORGANIC AND BIOMOLECULAR CHEMISTRY

SN - 1477-0520

IS - 28

ER -

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