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

Research output: Contribution to journalArticle

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

Fingerprint

Escherichia coli
Carrier Proteins
Fusion reactions
Proteins
Pseudomonas putida
Heat-Shock Proteins
Periplasmic Binding Proteins
Apoferritins
Ghrelin
Glutamate Decarboxylase
Putrescine
Spermidine
Inclusion Bodies
Granulocyte Colony-Stimulating Factor
Proteome
Epidermal Growth Factor
Solubility
Interleukin-2
Electrophoresis
Shielding

Keywords

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

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Genetics

Cite this

Transport proteins PotD and Crr of Escherichia coli, novel fusion partners for heterologous protein expression. / Han, Kyung Yeon; Seo, Hyuk Seong; Song, Jong A.; Ahn, Keum Young; Park, Jin Seung; Lee, Jeewon.

In: Biochimica et Biophysica Acta - Proteins and Proteomics, Vol. 1774, No. 12, 01.12.2007, p. 1536-1543.

Research output: Contribution to journalArticle

Han, Kyung Yeon ; Seo, Hyuk Seong ; Song, Jong A. ; Ahn, Keum Young ; Park, Jin Seung ; Lee, Jeewon. / Transport proteins PotD and Crr of Escherichia coli, novel fusion partners for heterologous protein expression. In: Biochimica et Biophysica Acta - Proteins and Proteomics. 2007 ; Vol. 1774, No. 12. pp. 1536-1543.
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AU - Seo, Hyuk Seong

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AU - Park, Jin Seung

AU - Lee, Jeewon

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AB - 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.

KW - Crr

KW - Escherichia coli BL21 proteome

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KW - Solubility enhancer

KW - Stress response

KW - Stress-responsive protein

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