Soluble Expression in Escherichia coli of Active Human Cyclic Nucleotide Phosphodiesterase Isoform 4B2 in Fusion with Maltose-Binding Protein

  • ZHU Sha
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • YANG Genqing
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University Laboratory Medicine Department, Third Affiliated Hospital of Xinxiang Medical College
  • YANG Xiaolan
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • ZHAO Yunsheng
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • LI Xiang
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • DENG Ping
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • XIE Yanling
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • GAN Zhiyong
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • LIU Yin
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • LI Zhirong
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • LIAO Juan
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • YU Ming’an
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University
  • LIAO Fei
    Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University

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  • Soluble Expression in<i>Escherichia coli</i>of Active Human Cyclic Nucleotide Phosphodiesterase Isoform 4B2 in Fusion with Maltose-Binding Protein

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Recombinant expression in Escherichia coli of human cyclic nucleotide phosphodiesterase 4B2 (hPDE4B2) fused to maltose-binding-protein (MBP-hPDE4B2) was investigated. hPDE4B2 DNA amplified via nested RT-PCR with total RNAs from U937 cells was ligated with pMAL-p2x. After induction at 18 °C for 16 h, soluble MBP-hPDE4B2 was produced in E. coli. MBP-hPDE4B2 after amylose-resin chromatography showed 35% homogeneity, and its Michaelis-Menten constant was 10±2 μM (n=3). Rolipram had a dissociation constant of 9±2 nM (n=2), and zinc ion was a potent inhibitor. Hence, MBP-hPDE4B2 was expressed in E. coli as a soluble active protein.

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