Abstract
Vibrio cholerae, the bacterium that causes the disease cholera, controls virulence factor production and biofilm development in response to two extracellular quorum-sensing molecules, called autoinducers. The strongest autoinducer, called CAI-1 (for cholera autoinducer-1), was previously identified as (S)-3-hydroxytridecan-4-one. Biosynthesis of CAI-1 requires the enzyme CqsA. Here, we determine the CqsA reaction mechanism, identify the CqsA substrates as (S)-2-aminobutyrate and decanoyl coenzyme A, and demonstrate that the product of the reaction is 3-aminotridecan-4-one, dubbed amino-CAI-1. CqsA produces amino-CAI-1 by a pyridoxal phosphate–dependent acyl-CoA transferase reaction. Amino-CAI-1 is converted to CAI-1 in a subsequent step via a CqsA-independent mechanism. Consistent with this, we find cells release ≥100 times more CAI-1 than amino-CAI-1. Nonetheless, V. cholerae responds to amino-CAI-1 as well as CAI-1, whereas other CAI-1 variants do not elicit a quorum-sensing response. Thus, both CAI-1 and amino-CAI-1 have potential as lead molecules in the development of an anticholera treatment.
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Acknowledgements
We thank the staff of the National Synchrotron Light Source beamline X29 for assistance with X-ray data collection; N. Ruiz and A. Arnaudo (Princeton University) for strains; and J. Kirsch for insightful discussions. This work was supported by the Howard Hughes Medical Institute; US National Institutes of Health grants AI054442, GM065859 and AI069326; US National Science Foundation grant MCB-0639855; and a Dr. Horst Witzel Fellowship from Cephalon Corporation (to M.E.B.).
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R.C.K., structural and spectroscopic analyses; M.E.B., chemistry; D.A.H. and W.-L.N., biology; W.L., mass spectrometry; P.D.J., crystallography. J.D.R., M.F.S., F.M.H. and B.L.B. provided guidance.
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Supplementary Figures 1–6, Supplementary Tables 1–5 and Supplementary Methods (PDF 1895 kb)
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Kelly, R., Bolitho, M., Higgins, D. et al. The Vibrio cholerae quorum-sensing autoinducer CAI-1: analysis of the biosynthetic enzyme CqsA. Nat Chem Biol 5, 891–895 (2009). https://doi.org/10.1038/nchembio.237
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DOI: https://doi.org/10.1038/nchembio.237
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