Journal article

Chemical probing suggests redox-regulation of the carbonic anhydrase activity of mycobacterial Rv1284

Lisa Nienaber, Elysia Cave-Freeman, Megan Cross, Lyndel Mason, Ulla-Maja Bailey, Parisa Amani, Rohan A Davis, Paul Taylor, Andreas Hofmann

FEBS JOURNAL | WILEY | Published : 2015

Abstract

UNLABELLED: The mycobacterial enzyme Rv1284 is a member of the β-carbonic anhydrase family that is considered essential for survival of the pathogen. The active site cavity of this dimeric protein is characterized by an exceptionally small volume and harbours a catalytic zinc ion coordinated by two cysteine and one histidine residue side chains. Using the natural products polycarpine and emodin as chemical probes in crystallographic experiments and stopped-flow enzyme assays, we report that the catalytic activity can be reversibly inhibited by oxidation. Oxidative conditions lead to the removal of one of the active site cysteine residues from the coordination sphere of the catalytic metal io..

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University of Melbourne Researchers

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Funding Acknowledgements

We gratefully acknowledge James Cameron (Griffith University) for help with the ICP-AES measurements, as well as Graeme Stevenson (Eskitis Institute) for useful discussions. Research in the laboratory of AH is funded by the National Health and Medical Research Council (NHMRC), the Australian Research Council (ARC) and the Rebecca L. Cooper Medical Research Foundation. RAD wishes to acknowledge the NHMRC (APP1024314) and ARC (LP120200339) for funding. Parts of this research were undertaken on the MX1 beamline at the Australian Synchrotron (Clayton, VIC, Australia).