Maltose binding protein facilitates high-level expression and functional purification of the chemokines RANTES and SDF-1α from Escherichia coli

Hee Jeong Cho, Young Lee, Rae Sung Chang, Moon Sun Hahm, Myung Kuk Kim, Young Bong Kim, Yu Kyoung Oh

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

The chemokines RANTES (regulated on activation, normal T cell expressed and secreted) and SDF-1α (stromal cell-derived factor-1α) are important regulators of leukocyte trafficking and homing. Chemokines form insoluble inclusion bodies when expressed in Escherichia coli (E. coli), resulting in low yields of soluble protein. We have developed a novel chemokine expression system that generates a high amount of soluble protein and uses a simple purification scheme. We cloned different types of RANTES and SDF-1α fused to either maltose binding protein (MBP) or glutathione-S-transferase (GST) and expressed the fusion proteins in E. coli under various conditions. We found that the yield of soluble chemokine is influenced by the type of fusion partner. Fusion to MBP resulted in a higher yield of total and soluble chemokine compared to GST. Under optimized conditions, the yield of soluble MBP-RANTES and MBP-SDF-1α was 2.5- and 4.5-fold higher than that of the corresponding GST-fusion protein, respectively. Recombinant chemokine fusion proteins exhibited specific binding activity to chemokine receptors. These results demonstrate that the use of MBP-fusion proteins may provide an approach to generating high yields of soluble and functional chemokines, such as RANTES and SDF-1α.

Original languageEnglish
Pages (from-to)37-45
Number of pages9
JournalProtein Expression and Purification
Volume60
Issue number1
DOIs
Publication statusPublished - 2008 Jul

Keywords

  • Chemokine
  • Escherichia coli
  • Fusion protein
  • RANTES
  • SDF-1α

ASJC Scopus subject areas

  • Biotechnology

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