Details

Autor(en) / Beteiligte

• Zhu, S.(Chongqing Medical Univ. (China))
• Yang, G
• Yang, X
• Zhao, Y
• Li, X
• Deng, P
• Xie, Y
• Gan, Z
• Liu, Y
• Li, Z
• Liao, J
• Yu, M
• Liao, F

Titel

Soluble expression in Escherichia coli of active human cyclic nucleotide phosphodiesterase isoform 4B2 in fusion with maltose-binding protein

Ist Teil von

• Bioscience, biotechnology, and biochemistry, 2009-04, Vol.73 (4), p.968-970

Ort / Verlag

Tokyo: Japan Society for Bioscience, Biotechnology, and Agrochemistry

Erscheinungsjahr

2009

Link zum Volltext

Quelle

Taylor & Francis Journals Auto-Holdings Collection

Beschreibungen/Notizen

• 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.