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Characterization and Inhibitor Screening of Plateau Zokor Lactate Dehydrogenase C4

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Abstract

Lactate dehydrogenase C4 (LDH-C4) is considered to be a target protein for the development of contraceptives. In this work, the characterization of plateau zokor LDH-C4 and the screening of a series of N-substituted oxamic acids as inhibitors against zokor LDH-C4 were reported. The cDNA of zokor LDH-C gene was cloned and expressed in Escherichia coli, from which the protein was purified and further characterized. The protein was a tetramer (LDH-C4) and thermally stable up to 62 °C with a K m of 63.9 μM for pyruvate and with optimal pH values of 7.95 and 10.1 for the forward and backward reactions respectively. Virtual and in vitro screening against zokor LDH-C4 revealed eight N-substituted oxamic acids with IC50s ranging from 198 to 2513 μM, higher than that of oxamic acid (150 μM) and (ethylamino)(oxo)acetic acid (59 μM). The inhibition potencies of N-substituted oxamic acids tested are in the micromolar range, and the increase in the length of substituting chain seems not to increase inhibition potency.

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Acknowledgments

We thank Dr. Min Yang for the language editing of this manuscript. Funds were provided by the National Science Foundation of China (31071700); the Youth Foundation of Sichuan province (2015JQO049); the scientific research fund of Sichuan Provincial Science and Technology Department (2015JY0232); Sichuan Provincial Health Department (130310); and the Fundamental Research Funds for the Central Universities, Southwest University for Nationalities (2015NYB13).

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    Authors and Affiliations

    1. College of Life Science and Technology, Southwest University for Nationalities, Chengdu, Sichuan, 610041, People’s Republic of China

      Qinghua He & Lin Huang

    2. School of Laboratory Medicine, Chengdu Medical College, Chengdu, Sichuan, 610500, People’s Republic of China

      Qinglian Zhang

    3. State Key Laboratory of Biotherapy and Cancer Center, West China Hospital, West China Medical School, Sichuan University, Sichuan, 610041, People’s Republic of China

      Jinhu Ma

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    1. Qinghua He
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    Correspondence to Qinghua He.

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    Qinghua He and Qinglian Zhang contributed equally to this work.

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    He, Q., Zhang, Q., Huang, L. et al. Characterization and Inhibitor Screening of Plateau Zokor Lactate Dehydrogenase C4. Appl Biochem Biotechnol 179, 927–937 (2016). https://doi.org/10.1007/s12010-016-2040-9

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    • DOI: https://doi.org/10.1007/s12010-016-2040-9

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