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Manganese superoxide dismutase from human pathogen Clostridium difficile

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Abstract

Clostridium difficile is a human pathogen that causes severe antibiotic-associated Clostridium difficile infection (CDI). Herein the MnSODcd from C. difficile was cloned, expressed in Escherichia Coli,and characterized by X-ray crystallography, UV/Vis and EPR spectroscopy, and activity assay, et al. The crystal structure of MnSODcd (2.32 Å) reveals a manganese coordination geometry of distorted trigonal bipyramidal, with His111, His197 and Asp193 providing the equatorial ligands and with His56 and a hydroxide or water forming the axial ligands. The catalytic activity of MnSODcd (8,600 U/mg) can be effectively inhibited by 2-methoxyestradiol with an IC50 of 75 μM. The affinity investigation between 2-methoxyestradiol and MnSODcd by ITC indicated a binding constant of 8.6 μM with enthalpy changes (ΔH = −4.08 ± 0.03 kcal/mol, ΔS = 9.53 ± 0.02 cal/mol/deg). An inhibitory mechanism of MnSODcd by 2-methoxyestradiol was probed and proposed based on molecular docking models and gel filtration analysis. The 2-methoxyestradiol may bind MnSODcd to interfere with the cross-linking between the two active sites of the dimer enzyme, compromising the SOD activity. These results provide valuable insight into the rational design of MnSODcd inhibitors for potential therapeutics for CDI.

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Acknowledgments

We sincerely thank Dr. Wei Tong from High Magnetic Field Laboratory of China Academy of Sciences, Prof. Jihu Su from University of Science and Technology, and Prof. Lindahl from Texas A&M University for EPR measurements and discussion. This work was financially supported partly by National Natural Science Foundation of China (No. 31270869, No. 21472027, No. 91013001), and the Ph.D. program of the Education Ministry of China (20100071110011).

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The authors declare that they have no competing financial interests.

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Authors and Affiliations

  1. Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China

    Wei Li

  2. Department of Chemistry, Fudan University, Shanghai, 200433, China

    Cheng Lei & Tianlei Ying

  3. State Key Laboratory of Quantum Optics and Quantum Optics Devices, Shanxi University, Taiyuan, 030006, China

    Hongfei Wang

  4. Department of Chemistry and Institutes of Biomedical Sciences, Fudan University, Shanghai, 200433, China

    Xiangshi Tan

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  1. Wei Li
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Correspondence to Xiangshi Tan.

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Handling Editor: F. Hollfelder.

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Li, W., Wang, H., Lei, C. et al. Manganese superoxide dismutase from human pathogen Clostridium difficile . Amino Acids 47, 987–995 (2015). https://doi.org/10.1007/s00726-015-1927-z

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