Abstract
LysW has been identified as a carrier protein in the lysine biosynthetic pathway that is active through the conversion of α-aminoadipate (AAA) to lysine. In this study, we found that the hyperthermophilic archaeon, Sulfolobus acidocaldarius, not only biosynthesizes lysine through LysW-mediated protection of AAA but also uses LysW to protect the amino group of glutamate in arginine biosynthesis. In this archaeon, after LysW modification, AAA and glutamate are converted to lysine and ornithine, respectively, by a single set of enzymes with dual functions. The crystal structure of ArgX, the enzyme responsible for modification and protection of the amino moiety of glutamate with LysW, was determined in complex with LysW. Structural comparison and enzymatic characterization using Sulfolobus LysX, Sulfolobus ArgX and Thermus LysX identify the amino acid motif responsible for substrate discrimination between AAA and glutamate. Phylogenetic analysis reveals that gene duplication events at different stages of evolution led to ArgX and LysX.
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Acknowledgements
This work was supported in part by a grant-in-aid for scientific research from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (grant nos. 21380057 and 24228001 to M.N.), Nagase Science and Technology Foundation to M.N., and from the Asahi Glass Foundation to M.N.
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Research planning and supervision were by T.T., T.K. and M.N.; biochemical experiments were by T.O., K.T., A.H., A.Y. and S. Kosono; phylogenetic analysis was by H.N.; gene knockout and replacement of S. acidocaldarius was by K.L. and S.-V.A.; LC-MS/MS and MALDI-TOF MS was by H.T., R. Mineki, T.F. and C.N.; StArgX was purified by T.O., A.H., A.Y., R. Masui and S. Kuramitsu; Crystallographic analysis was by T.O., A.H. and T.T.; and the manuscript was written by T.O., T.T., H.N., S.-V.A. and M.N.
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Ouchi, T., Tomita, T., Horie, A. et al. Lysine and arginine biosyntheses mediated by a common carrier protein in Sulfolobus. Nat Chem Biol 9, 277–283 (2013). https://doi.org/10.1038/nchembio.1200
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DOI: https://doi.org/10.1038/nchembio.1200
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