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Title: Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity

Abstract

APS kinase from Thiobacillus denitrificans contains an inactive N-terminal ATP sulfurylase domain. The structure presented unveils the first hexameric assembly for an APS kinase, and reveals that structural changes in the N-terminal domain disrupt the ATP sulfurylase active site thus prohibiting activity. The Tbd-0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 Å resolution X-ray crystal structure reported here revealed a hexameric assembly with D{sub 3} symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.

Authors:
 [1];  [2];  [1];  [3]
  1. Department of Chemistry, University of California, One Shields Avenue, Davis, CA 95616 (United States)
  2. Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616 (United States)
  3. (United States)
Publication Date:
OSTI Identifier:
22351193
Resource Type:
Journal Article
Journal Name:
Acta Crystallographica. Section D: Biological Crystallography
Additional Journal Information:
Journal Volume: 65; Journal Issue: Pt 10; Other Information: PMCID: PMC2756168; PMID: 19770499; PUBLISHER-ID: hm5075; OAI: oai:pubmedcentral.nih.gov:2756168; Copyright (c) International Union of Crystallography 2009; Country of input: International Atomic Energy Agency (IAEA); Journal ID: ISSN 0907-4449
Country of Publication:
Denmark
Language:
English
Subject:
75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND SUPERFLUIDITY; CRYSTAL STRUCTURE; RESOLUTION; SYMMETRY

Citation Formats

Gay, Sean C., Segel, Irwin H., Fisher, Andrew J., E-mail: fisher@chem.ucdavis.edu, and Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity. Denmark: N. p., 2009. Web. doi:10.1107/S0907444909026547.
Gay, Sean C., Segel, Irwin H., Fisher, Andrew J., E-mail: fisher@chem.ucdavis.edu, & Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616. Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity. Denmark. doi:10.1107/S0907444909026547.
Gay, Sean C., Segel, Irwin H., Fisher, Andrew J., E-mail: fisher@chem.ucdavis.edu, and Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616. Thu . "Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity". Denmark. doi:10.1107/S0907444909026547.
@article{osti_22351193,
title = {Structure of the two-domain hexameric APS kinase from Thiobacillus denitrificans: structural basis for the absence of ATP sulfurylase activity},
author = {Gay, Sean C. and Segel, Irwin H. and Fisher, Andrew J., E-mail: fisher@chem.ucdavis.edu and Section of Molecular and Cellular Biology, University of California, One Shields Avenue, Davis, CA 95616},
abstractNote = {APS kinase from Thiobacillus denitrificans contains an inactive N-terminal ATP sulfurylase domain. The structure presented unveils the first hexameric assembly for an APS kinase, and reveals that structural changes in the N-terminal domain disrupt the ATP sulfurylase active site thus prohibiting activity. The Tbd-0210 gene of the chemolithotrophic bacterium Thiobacillus denitrificans is annotated to encode a 60.5 kDa bifunctional enzyme with ATP sulfurylase and APS kinase activity. This putative bifunctional enzyme was cloned, expressed and structurally characterized. The 2.95 Å resolution X-ray crystal structure reported here revealed a hexameric assembly with D{sub 3} symmetry. Each subunit contains a large N-terminal sulfurylase-like domain and a C-terminal APS kinase domain reminiscent of the two-domain fungal ATP sulfurylases of Penicillium chrysogenum and Saccharomyces cerevisiae, which also exhibit a hexameric assembly. However, the T. denitrificans enzyme exhibits numerous structural and sequence differences in the N-terminal domain that render it inactive with respect to ATP sulfurylase activity. Surprisingly, the C-terminal domain does indeed display APS kinase activity, indicating that this gene product is a true APS kinase. Therefore, these results provide the first structural insights into a unique hexameric APS kinase that contains a nonfunctional ATP sulfurylase-like domain of unknown function.},
doi = {10.1107/S0907444909026547},
journal = {Acta Crystallographica. Section D: Biological Crystallography},
issn = {0907-4449},
number = Pt 10,
volume = 65,
place = {Denmark},
year = {2009},
month = {10}
}