Transport proteins PotD and Crr of Escherichia coli, novel fusion partners for heterologous protein expression

Kyung Yeon Han, Hyuk Seong Seo, Jong A. Song, Keum Young Ahn, Jin Seung Park, Jeewon Lee

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17 Citations (Scopus)

Abstract

The Escherichia coli proteome response to the stressor GdnHCl was analyzed through 2-dimensional gel electrophoresis (2-DE). We identified PotD (spermidine/putrescine-binding periplasmic protein) and Crr [glucose-specific phosphotransferase (PTS) enzyme IIA component] as a stress-responsive protein. Even under a stress situation where the total number of soluble proteins decreased by about 10%, 3.5- and 2.2-fold increase was observed in the synthesis of PotD and Crr, respectively. As fusion partners, PotD and Crr dramatically increased the solubility of many aggregation-prone heterologous proteins [e.g. human minipro-insulin (mp-INS), human epidermal growth factor (EGF), human prepro-ghrelin (ppGRN), human interleukin-2(hIL-2), human activation induced cytidine deaminase (AID), human glutamate decarboxylase (GAD448-585), Pseudomonas putida cutinase (CUT), human ferritin light chain (hFTN-L), human granulocyte colony-stimulating factor (G-CSF), and cold autoinflammatory syndrome1 protein (NALP3) Nacht domain (NACHT)] in the E. coli cytoplasm. Presumably PotD and Crr were very effective in shielding interactive surfaces of heterologous proteins associated with non-specific protein-protein interactions leading to the formation of inclusion bodies most likely due to intrinsic high folding efficiency, chaperone-like activity, or a combination of both factors. Both the stress-induced proteins were well suited for the production of a biologically active fusion mutant of P. putida cutinase that can be expected to be of biotechnological and commercial interest.

Original languageEnglish
Pages (from-to)1536-1543
Number of pages8
JournalBiochimica et Biophysica Acta - Proteins and Proteomics
Volume1774
Issue number12
DOIs
Publication statusPublished - 2007 Dec 1

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Keywords

  • Crr
  • Escherichia coli BL21 proteome
  • PotD
  • Solubility enhancer
  • Stress response
  • Stress-responsive protein

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics

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