Zhu Sha, Yang Genqing, Yang Xiaolan, Zhao Yunsheng, Li Xiang, Deng Ping, Xie Yanling, Gan Zhiyong, Liu Yin, Li Zhirong, Liao Juan, Yu Ming'an, Liao Fei
Chongqing Key Laboratory of Biochemical and Molecular Pharmacology, Chongqing Medical University, Chongqing 400016, China.
Biosci Biotechnol Biochem. 2009 Apr 23;73(4):968-70. doi: 10.1271/bbb.90131. Epub 2009 Apr 7.
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 degrees 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 microM (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.
