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Interaction between DAHP synthase and chorismate mutase endows new regulation on DAHP synthase activity in Corynebacterium glutamicum

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Abstract

Previous research on Corynebacterium glutamicum revealed that 3-deoxy-d-arabino-heptulosonate 7-phosphate synthase (DSCg, formerly DS2098) interacts with chorismate mutase (CMCg, formerly CM0819). In this study, we investigated the interaction by means of structure-guided mutation and enzymatic assays. Our results show that the interaction imparted a new mechanism for regulation of DAHP activity: In the absence of CMCg, DSCg activity was not regulated by prephenate, whereas in the presence of CMCg, prephenate markedly inhibited DSCg activity. Prephenate competed with the substrate phosphoenolpyruvate, and the inhibition constant (K i) was determined to be 0.945 mM. Modeling based on the structure of the complex formed between DAHP synthase and chorismate mutase of Mycobacterium tuberculosis predicted the interaction surfaces of the putative DSCg-CMCg complex. The amino acid residues and structural domains that contributed to the interaction surfaces were experimentally identified to be the 212SPAGARYE219 sequence of DSCg and the 60SGGTR64 loop and C-terminus (97RGKLG101) of CMCg.

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Acknowledgments

This work was supported by a grant from the Ministry of Science and Technology of China (973-project, grant no. 2012CB721104).

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Authors and Affiliations

  1. State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Pan-Pan Li, Yi-Ming Liu, Chang Liu & Shuang-Jiang Liu

  2. Institute of Biophysics, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    De-Feng Li

  3. Network Information Center, Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, People’s Republic of China

    Di Liu

  4. Environmental Microbiology and Biotechnology Research Center, Institute of Microbiology, Chinese Academy of Sciences, Beichen West Road No. 1, Chaoyang District, Beijing, 100101, People’s Republic of China

    Yi-Ming Liu & Shuang-Jiang Liu

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  1. Pan-Pan Li
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  2. De-Feng Li
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Correspondence to Shuang-Jiang Liu.

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Li, PP., Li, DF., Liu, D. et al. Interaction between DAHP synthase and chorismate mutase endows new regulation on DAHP synthase activity in Corynebacterium glutamicum . Appl Microbiol Biotechnol 97, 10373–10380 (2013). https://doi.org/10.1007/s00253-013-4806-0

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  • DOI: https://doi.org/10.1007/s00253-013-4806-0

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