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Title: Functional Profiling and Crystal Structures of Isothiocyanate Hydrolases Found in Gut-Associated and Plant-Pathogenic Bacteria

Abstract

Isothiocyanates (ITCs) are produced by cruciferous plants to protect them against herbivores and infection by microbes. These compounds are of particular interest due to their antimicrobial and anticarcinogenic properties. The breakdown of ITCs in nature is catalyzed by isothiocyanate hydrolases (ITCases), a novel family within the metallo-β-lactamase (MBL)-fold superfamily of proteins. saxA genes that code for ITCases are particularly widespread in insect- and plant-associated bacteria. Enzymatic characterization of seven phylogenetically related but distinct ITCases revealed similar activities on six selected ITCs, suggesting that phylogenetic diversity does not determine the substrate specificity of ITCases. X-ray crystallography studies of two ITCases sharing 42% amino acid sequence identity revealed a highly conserved tertiary structure. Notable features of ITCases include a hydrophobic active site with two Zn2+ ions coordinating water/hydroxide and a flexible cap that is implicated in substrate recognition and covers the active site. Furthermore, this report reveals the function and structure of the previously uncharacterized family of isothiocyanate hydrolases within the otherwise relatively well-studied superfamily of metallo-β-lactamases.

Authors:
 [1];  [2];  [2]; ORCiD logo [3];  [4]
  1. Radboud Univ., Nijmegen (The Netherlands)
  2. Argonne National Lab. (ANL), Lemont, IL (United States); Univ. of Chicago, Chicago, IL (United States)
  3. Radboud Univ., Nijmegen (The Netherlands); Soehngen Institute for Anaerobic Microbiology, Nijmegen (The Netherlands)
  4. Univ. of Tartu, Tartu (Estonia)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER); Netherlands Organisation for Scientific Research (NWO)
OSTI Identifier:
1467602
Grant/Contract Number:  
AC02-06CH11357
Resource Type:
Accepted Manuscript
Journal Name:
Applied and Environmental Microbiology
Additional Journal Information:
Journal Volume: 84; Journal Issue: 14; Journal ID: ISSN 0099-2240
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; SaxA; metallo-β-lactamase superfamily; multisubstrate enzyme assay; X-ray crystallography; isothiocyanate hydrolase

Citation Formats

van den Bosch, Tijs J. M., Tan, Kemin, Joachimiak, Andrzej, Welte, Cornelia U., and Kivisaar, Maia. Functional Profiling and Crystal Structures of Isothiocyanate Hydrolases Found in Gut-Associated and Plant-Pathogenic Bacteria. United States: N. p., 2018. Web. doi:10.1128/AEM.00478-18.
van den Bosch, Tijs J. M., Tan, Kemin, Joachimiak, Andrzej, Welte, Cornelia U., & Kivisaar, Maia. Functional Profiling and Crystal Structures of Isothiocyanate Hydrolases Found in Gut-Associated and Plant-Pathogenic Bacteria. United States. https://doi.org/10.1128/AEM.00478-18
van den Bosch, Tijs J. M., Tan, Kemin, Joachimiak, Andrzej, Welte, Cornelia U., and Kivisaar, Maia. Mon . "Functional Profiling and Crystal Structures of Isothiocyanate Hydrolases Found in Gut-Associated and Plant-Pathogenic Bacteria". United States. https://doi.org/10.1128/AEM.00478-18. https://www.osti.gov/servlets/purl/1467602.
@article{osti_1467602,
title = {Functional Profiling and Crystal Structures of Isothiocyanate Hydrolases Found in Gut-Associated and Plant-Pathogenic Bacteria},
author = {van den Bosch, Tijs J. M. and Tan, Kemin and Joachimiak, Andrzej and Welte, Cornelia U. and Kivisaar, Maia},
abstractNote = {Isothiocyanates (ITCs) are produced by cruciferous plants to protect them against herbivores and infection by microbes. These compounds are of particular interest due to their antimicrobial and anticarcinogenic properties. The breakdown of ITCs in nature is catalyzed by isothiocyanate hydrolases (ITCases), a novel family within the metallo-β-lactamase (MBL)-fold superfamily of proteins. saxA genes that code for ITCases are particularly widespread in insect- and plant-associated bacteria. Enzymatic characterization of seven phylogenetically related but distinct ITCases revealed similar activities on six selected ITCs, suggesting that phylogenetic diversity does not determine the substrate specificity of ITCases. X-ray crystallography studies of two ITCases sharing 42% amino acid sequence identity revealed a highly conserved tertiary structure. Notable features of ITCases include a hydrophobic active site with two Zn2+ ions coordinating water/hydroxide and a flexible cap that is implicated in substrate recognition and covers the active site. Furthermore, this report reveals the function and structure of the previously uncharacterized family of isothiocyanate hydrolases within the otherwise relatively well-studied superfamily of metallo-β-lactamases.},
doi = {10.1128/AEM.00478-18},
journal = {Applied and Environmental Microbiology},
number = 14,
volume = 84,
place = {United States},
year = {Mon Jul 02 00:00:00 EDT 2018},
month = {Mon Jul 02 00:00:00 EDT 2018}
}

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Works referencing / citing this record:

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