Affordable Access

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

Authors
  • 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
Type
Published Article
Journal
Bioscience Biotechnology and Biochemistry
Publisher
Informa UK (Taylor & Francis)
Publication Date
Apr 23, 2009
Volume
73
Issue
4
Pages
968–970
Identifiers
PMID: 19352009
Source
Medline
License
Unknown

Abstract

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.

Report this publication

Statistics

Seen <100 times