Journal article
Structural and kinetic insights into the stereospecific oxidation of R-2,3-dihydroxypropanesulfonate by DHPS-3-dehydrogenase from Cupriavidus pinatubonensis
Laura Burchill, Arashdeep Kaur, Artur Nastasovici, Mihwa Lee, Spencer J Williams
Chemical Science | The Royal Society of Chemistry | Published : 2024
DOI: 10.1039/d4sc05114a
Abstract
2,3-Dihydroxypropanesulfonate (DHPS) and sulfolactate (SL) are environmentally important organosulfur compounds that play key roles as metabolic currencies in the sulfur cycle. Despite their prevalence, the pathways governing DHPS and SL production remain poorly understood. Here, we study DHPS-3-dehydrogenase from Cupriavidus pinatubonensis (CpHpsN), a bacterium capable of utilizing DHPS as a sole carbon source. Kinetic analysis of CpHpsN reveals a strict preference for R-DHPS, catalyzing its 4-electron oxidation to R-SL, with high specificity for NAD+ over NADP+. The 3D structure of CpHpsN in complex with Zn2+, NADH and R-SL, elucidated through X-ray crystallography, reveals a fold akin to ..
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Grants
Awarded by Australian Research Council
Funding Acknowledgements
This work was supported by the Australian Research Council (DP210100233, DP230102668, DP240100126), A. K. is supported by a Norma Hilda Schuster Scholarship. L. B. is supported by an Elizabeth Vernon Puzey Postdoctoral Fellowship and Thomas Davies research grant (Australian Academy of Sciences). M. L. is a recipient of Kaye Merlin Brutton Bequest Funding (University of Melbourne). Aspects of this research were undertaken on the Macromolecular Crystallography beamlines at the Australian Synchrotron (Victoria, Australia), part of ANSTO, and made use of the Australian Cancer Research Foundation (ACRF) detector. We thank the Australian Synchrotron beamline staff for their professional support. We acknowledge use of the Melbourne Mass Spectrometry and Proteomics Facility, and the Melbourne Protein Characterisation Platform at The University of Melbourne. We thank Prof. Megan Maher (Bio21 Institute, University of Melbourne) for helpful discussion and advice on inorganic and crystallographic aspects of the manuscript, and generous access to research infrastructure.